Animal nutrition strategies and options to reduce the use of antimicrobials in animal production

Antimicrobial resistance is a global and increasing threat. Stewardship campaigns have been established, and policies implemented, to safeguard the appropriate use of antimicrobials in humans, animals and plants. Restrictions on the use of antimicrobials in animal production are on the agenda worldwide. Producers are investing in measures, involving biosecurity, genetics, health care, farm management, animal welfare and nutrition, to prevent diseases and minimize the use of antimicrobials. Young animals (piglets, broiler chickens and calves) are particularly susceptible to diseases and disorders, and the use of antimicrobials on these animals is therefore relatively high. Functional nutrition to promote animal health is one of the tools available to decrease the need for antimicrobials in animal production. Nutrition affects the critical functions required for host defence and disease resistance. Animal nutrition strategies should therefore aim to support these host defence systems and reduce the risk of the presence in feed and water of potentially harmful substances, such as mycotoxins, anti-nutritional factors and pathogenic bacteria and other microbes. General dietary measures to promote gastrointestinal tract (GIT) health include, for example, the functional use of dietary fibres to stimulate gastrointestinal secretions and motility, lowering the protein content to avoid excessive fermentation of protein in the hindgut, and selective use of a combination of feed additives and feed ingredients to stabilize the intestinal microbiota and support mucosal barrier function. In addition, the use of organic acids may contribute to feed and water safety. This knowledge, used to establish best practices in animal nutrition, could allow the adoption of strategies to reduce the need for antimicrobials and contain antimicrobial resistance

Efficacy of l-glutamic acid, N,N-diacetic acid to improve the dietary trace mineral bioavailability in broilers

Trace minerals are commonly supplemented in the diets of farmed animals in levels exceeding biological requirements, resulting in extensive fecal excretion and environmental losses. Chelation of trace metal supplements with ethylenediaminetetraacetic acid (EDTA) can mitigate the effects of dietary antagonists by preserving the solubility of trace minerals. Lack of EDTA biodegradability, however, is of environmental concern. l-Glutamic acid, N,N-diacetic acid (GLDA) is a readily biodegradable chelating agent that could be used as a suitable alternative to EDTA. The latter was tested in sequential dose-response experiments in broiler chickens. Study 1 compared the effect of EDTA and GLDA in broilers on supplemental zinc availability at three levels of added zinc (5, 10, and 20 ppm) fed alone or in combination with molar amounts of GLDA or EDTA equivalent to chelate the added zinc, including negative (no supplemental zinc) and positive (80 ppm added zinc) control treatments. Study 2 quantified the effect of GLDA on the availability of native trace mineral feed content in a basal diet containing no supplemental minerals and supplemented with three levels of GLDA (54, 108, and 216 ppm). In study 1, serum and tibia Zn clearly responded to the increasing doses of dietary zinc with a significant response to the presence of EDTA and GLDA (P < 0.05). These results are also indicative of the equivalent nutritional properties between GLDA and EDTA. In study 2, zinc levels in serum and tibia were also increased with the addition of GLDA to a basal diet lacking supplemental trace minerals, where serum zinc levels were 60% higher at the 216 ppm inclusion level. Similar to the reported effects of EDTA, these studies demonstrate that dietary GLDA may have enhanced zinc solubility in the gastrointestinal tract and subsequently enhanced availability for absorption, resulting in improved nutritional zinc status in zinc-deficient diets. As such, GLDA can be an effective nutritional tool to reduce supplemental zinc levels in broiler diets, thereby maintaining health and performance while reducing the environmental footprint of food-producing animals.</p

The availability of drug by liposomal drug delivery : Individual kinetics and tissue distribution of encapsulated and released drug in mice after administration of PEGylated liposomal prednisolone phosphate

Lately, the usefulness of liposomal drug delivery systems has been debated. To better understand the underlying pharmacokinetics of the targeted drug delivery by liposomes, individual encapsulated and non-encapsulated drug concentrations in blood, tumor, liver, spleen and kidneys were quantified after i.v. administration of liposomal prednisolone phosphate in mice. Kinetic analysis shows that the tumor influx of encapsulated drug is not dominant compared to the uptake by the other tissues. Further, from a quantitative point of view, the availability of non-encapsulated drug in the tumor tissue after liposomal delivery is not pronounced as compared to the other tissues studied. However, drug release in the tumor seems more extended than in the other tissues and the non-encapsulated drug concentration decreases more slowly in the tumor than in the liver and spleen. The spleen shows a high affinity for the uptake of encapsulated drug as well as the release of drug from the liposomes. Subsequently, released drug in the spleen, and possibly also in other tissues, is probably quickly redistributed towards the blood and other tissues. This also impairs the drug delivery effect of the liposomes. In contrast to the released drug in the central circulation, liver and spleen, the released drug concentration in the tumor remains at a fairly constant level likely due to the extended release kinetics from the liposomes. These extended release characteristics in the tumor most probably contribute to the beneficial effect. Nevertheless, it should be noted that larger released drug concentrations are formed in healthy tissues

The availability of drug by liposomal drug delivery: Individual kinetics and tissue distribution of encapsulated and released drug in mice after administration of PEGylated liposomal prednisolone phosphate

Lately, the usefulness of liposomal drug delivery systems has been debated. To better understand the underlying pharmacokinetics of the targeted drug delivery by liposomes, individual encapsulated and non-encapsulated drug concentrations in blood, tumor, liver, spleen and kidneys were quantified after i.v. administration of liposomal prednisolone phosphate in mice. Kinetic analysis shows that the tumor influx of encapsulated drug is not dominant compared to the uptake by the other tissues. Further, from a quantitative point of view, the availability of non-encapsulated drug in the tumor tissue after liposomal delivery is not pronounced as compared to the other tissues studied. However, drug release in the tumor seems more extended than in the other tissues and the non-encapsulated drug concentration decreases more slowly in the tumor than in the liver and spleen. The spleen shows a high affinity for the uptake of encapsulated drug as well as the release of drug from the liposomes. Subsequently, released drug in the spleen, and possibly also in other tissues, is probably quickly redistributed towards the blood and other tissues. This also impairs the drug delivery effect of the liposomes. In contrast to the released drug in the central circulation, liver and spleen, the released drug concentration in the tumor remains at a fairly constant level likely due to the extended release kinetics from the liposomes. These extended release characteristics in the tumor most probably contribute to the beneficial effect. Nevertheless, it should be noted that larger released drug concentrations are formed in healthy tissues

A reactive center loop-based prediction platform to enhance the design of therapeutic SERPINs

Serine proteases are essential for many physiological processes and require tight regulation by serine protease inhibitors (SERPINs). A disturbed SERPIN-protease balance may result in disease. The reactive center loop (RCL) contains an enzymatic cleavage site between the P1 through P1' residues that controls SERPIN specificity. This RCL can be modified to improve SERPIN function; however, a lack of insight into sequence-function relationships limits SERPIN development. This is complicated by more than 25 billion mutants needed to screen the entire P4 to P4' region. Here, we developed a platform to predict the effects of RCL mutagenesis by using α1-antitrypsin as a model SERPIN. We generated variants for each of the residues in P4 to P4' region, mutating them into each of the 20 naturally occurring amino acids. Subsequently, we profiled the reactivity of the resulting 160 variants against seven proteases involved in coagulation. These profiles formed the basis of an in silico prediction platform for SERPIN inhibitory behavior with combined P4 to P4' RCL mutations, which were validated experimentally. This prediction platform accurately predicted SERPIN behavior against five out of the seven screened proteases, one of which was activated protein C (APC). Using these findings, a next-generation APC-inhibiting α1-antitrypsin variant was designed (KMPR/RIRA; / indicates the cleavage site). This variant attenuates blood loss in an in vivo hemophilia A model at a lower dosage than the previously developed variant AIKR/KIPP because of improved potency and specificity. We propose that this SERPIN-based RCL mutagenesis approach improves our understanding of SERPIN behavior and will facilitate the design of therapeutic SERPINs

A reactive center loop-based prediction platform to enhance the design of therapeutic SERPINs

Serine proteases are essential for many physiological processes and require tight regulation by serine protease inhibitors (SERPINs). A disturbed SERPIN-protease balance may result in disease. The reactive center loop (RCL) contains an enzymatic cleavage site between the P1 through P1' residues that controls SERPIN specificity. This RCL can be modified to improve SERPIN function; however, a lack of insight into sequence-function relationships limits SERPIN development. This is complicated by more than 25 billion mutants needed to screen the entire P4 to P4' region. Here, we developed a platform to predict the effects of RCL mutagenesis by using α1-antitrypsin as a model SERPIN. We generated variants for each of the residues in P4 to P4' region, mutating them into each of the 20 naturally occurring amino acids. Subsequently, we profiled the reactivity of the resulting 160 variants against seven proteases involved in coagulation. These profiles formed the basis of an in silico prediction platform for SERPIN inhibitory behavior with combined P4 to P4' RCL mutations, which were validated experimentally. This prediction platform accurately predicted SERPIN behavior against five out of the seven screened proteases, one of which was activated protein C (APC). Using these findings, a next-generation APC-inhibiting α1-antitrypsin variant was designed (KMPR/RIRA; / indicates the cleavage site). This variant attenuates blood loss in an in vivo hemophilia A model at a lower dosage than the previously developed variant AIKR/KIPP because of improved potency and specificity. We propose that this SERPIN-based RCL mutagenesis approach improves our understanding of SERPIN behavior and will facilitate the design of therapeutic SERPINs

A reactive center loop-based prediction platform to enhance the design of therapeutic SERPINs

Serine proteases are essential for many physiological processes and require tight regulation by serine protease inhibitors (SERPINs). A disturbed SERPIN-protease balance may result in disease. The reactive center loop (RCL) contains an enzymatic cleavage site between the P1 through P1' residues that controls SERPIN specificity. This RCL can be modified to improve SERPIN function; however, a lack of insight into sequence-function relationships limits SERPIN development. This is complicated by more than 25 billion mutants needed to screen the entire P4 to P4' region. Here, we developed a platform to predict the effects of RCL mutagenesis by using α1-antitrypsin as a model SERPIN. We generated variants for each of the residues in P4 to P4' region, mutating them into each of the 20 naturally occurring amino acids. Subsequently, we profiled the reactivity of the resulting 160 variants against seven proteases involved in coagulation. These profiles formed the basis of an in silico prediction platform for SERPIN inhibitory behavior with combined P4 to P4' RCL mutations, which were validated experimentally. This prediction platform accurately predicted SERPIN behavior against five out of the seven screened proteases, one of which was activated protein C (APC). Using these findings, a next-generation APC-inhibiting α1-antitrypsin variant was designed (KMPR/RIRA; / indicates the cleavage site). This variant attenuates blood loss in an in vivo hemophilia A model at a lower dosage than the previously developed variant AIKR/KIPP because of improved potency and specificity. We propose that this SERPIN-based RCL mutagenesis approach improves our understanding of SERPIN behavior and will facilitate the design of therapeutic SERPINs

Effects of a feed additive blend on broilers challenged with heat stress

We evaluated a blend of medium-chain fatty acids (MCFA), organic acids, and a polyphenol antioxidant on gut integrity. Eighty Ross Broilers were exposed to 20–22°C (control – normothermic) or to 35–39.5°C (heat stress) for eight hours a day for a period of 1 or 5 days. Birds were fed a standard diet, or a diet supplemented with the test blend. Thereafter, birds were euthanized, and intestinal sections were excised for morphological, morphometric and gene expression analyses. Blood samples were collected for glucose-6-phosphate dehydrogenase (G6PD), glutathione peroxidase (GSH-Px) activity and trolox equivalent antioxidant capacity (TEAC) determination. Heart and liver tissues were used to quantify the expression of heat shock proteins 60 and 70 (HSP60 and HSP70, respectively) and inhibitor of kappa light chain gene enhancer in B cells alpha (IKBA). The jejunum was the most sensitive intestinal section, where heat stress modulated the expression of HSP70, of the inflammatory markers IKBA, interleukin 8 (IL-8), interferon gamma (IFNγ), and toll-like receptor 4 (TLR4). Moreover, expression of tight junctions (CLDN1, ZO1 and ZO2) and nutrient transporters (PEPT1 and EAAT3) was modulated especially in the jejunum. In conclusion, the feed additive blend protected intestines during heat stress from the decrease in villus height and crypt depth, and from the increase in villus width. Especially in the jejunum, heat stress played an important role by modulating oxidative stress and inflammation, impairing gut integrity and nutrient transport, and such deleterious effects were alleviated by the feed additive blend