Withdrawal Of Antibiotic Growth Promoters Research Articles

Antibiotics in avian care and husbandry-status and alternative antimicrobials

Abstract Undoubtedly, the discovery of antibiotics was one of the greatest milestones in the treatment of human and animal diseases. Due to their over-use mainly as antibiotic growth promoters (AGP) in livestock farming, antimicrobial resistance has been reported with increasing intensity, especially in the last decades. In order to reduce the scale of this phenomenon, initially in the Scandinavian countries and then throughout the entire European Union, a total ban on the use of AGP was introduced, moreover, a significant limitation in the use of these feed additives is now observed almost all over the world. The withdrawal of AGP from widespread use has prompted investigators to search for alternative strategies to maintain and stabilize the composition of the gut microbiota. These strategies include substances that are used in an attempt to stimulate the growth and activity of symbiotic bacteria living in the digestive tract of animals, as well as living microorganisms capable of colonizing the host’s gastrointestinal tract, which can positively affect the composition of the intestinal microbiota by exerting a number of pro-health effects, i.e., prebiotics and probiotics, respectively. In this review we also focused on plants/herbs derived products that are collectively known as phytobiotic.

Antioxidant and Antimicrobial Properties of Selected Phytogenics for Sustainable Poultry Production

The use of antibiotic growth promoters (AGP) in poultry production not only promotes the emergence of pathogenic multi-drug resistant bacteria, but it also compromises product quality, threatens animal and human health, and pollutes the environment. However, the complete withdrawal of AGP without alternatives could result in uncontrollable disease outbreaks that would jeopardize large-scale poultry intensification. Thus, the use of phytogenic products as potential alternatives to in-feed AGP has attracted worldwide research interest. These phytogenic products contain numerous biologically active substances with antioxidant and antimicrobial activities that can enhance poultry health, growth performance, and meat quality characteristics. In addition, the incorporation of phytogenic products as feed additives in poultry diets could result in the production of high-quality, drug-free, and organic poultry products that are safe for human consumption. Thus, this review examines the current evidence on the antioxidant and antimicrobial properties of a selection of phytogenic products, their effects on nutrient utilization, and physiological and meat quality parameters in poultry. The paper also reviews the factors that could limit the utilization of phytogenic products in poultry nutrition and proposes solutions that can deliver efficient and sustainable poultry production systems for global food and nutrition security.

Withdrawal of antibiotic growth promoters in China and its impact on the foodborne pathogen Campylobacter coli of swine origin.

Antibiotic growth promoters (AGPs) have been used as feed additives to improve feed efficiency in food animals for more than six decades. However, the wide use of AGPs has led to the emergence of antibiotic-resistant pathogens of animal origin, posing a significant threat to food safety and public health. China prohibited the addition of AGPs to animal feed from July 2020. The impacts caused by the withdrawal of AGPs on the prevalence and antibiotic resistance of foodborne pathogens have not been illustrated. Here, a total of 471 strains of Campylobacter were isolated from pigs from three pig farms and two slaughterhouses in Sichuan Province for 4 consecutive years (2018–2021), including 2 years before and 2 years after the ban on AGPs in China. The isolation rate of Campylobacter had a slight increase after prohibiting the addition of AGPs to the feed. Contrary to what we expected, the antibiotic susceptibility test and WGS data showed that the antibiotic resistance to gentamicin and florfenicol and the abundance of virulence genes increased significantly after the ban of AGPs. Comparison of the isolates of swine origin with isolates of human origin indicated the potential of antibiotic-resistant Campylobacter transmission from pigs to humans. These data suggested that phasing out AGPs may lead to increased use of therapeutic antimicrobials, promoting the prevalence and transmission of both antibiotic resistance and virulence genes.

Immunization with Pooled Antigens for Clostridium perfringens Conferred Partial Protection against Experimental Necrotic Enteritis in Broiler Chickens.

Necrotic enteritis (NE) is a multifactorial and important enteric infectious disease etiologically caused by pathogenic C. perfringens infection, accounting for the estimated loss of around USD 6 billion in the global poultry industry. The increasing incidence of NE was found to be associated with the voluntary reduction or withdrawal of antibiotic growth promoters from animal feed during recent years. Therefore, the development of effective vaccines specific to NE assumes a priority for the poultry industry. This study aimed to identify the potential C. perfringens proteins as vaccine targets for NE. Three recombinant C. perfringens proteins targeting five antigens were prepared: two chimeric proteins (alpha-toxin and NetB, fructose-1,6-bisphosphate aldolase (FBA) and a zinc metalloprotease (Zm)), and one single collagen adhesion protein (Cna). Their protection efficacies were evaluated with a potent challenge model of Eimeria maxima/C. perfringens dual infections using a netB+tpeL+ C. perfringens strain. Young chicks were immunized twice subcutaneously with adjuvanted C. perfringens proteins on Days 4 and 15. At six days after the second immunization, the chickens immunized with Cna, FBA, and Zm antigens, and alpha-toxin had much higher serum antibody titers than unvaccinated controls prior to the challenge. Following the challenge, the pooled antigen-immunized group demonstrated no mortality and the least lesion scores against virulent challenge. The results indicate that the immunization with multicomponent antigens, including C. perfringens housekeeping protein Cna, may confer partial protection.

Estimation of productive losses caused by withdrawal of antibiotic growth promoter from pig diets – Meta-analysis

ABSTRACT: This study was designed to simulate productive and economic losses due to the withdrawal of antibiotic growth promoters (AGP) from pig diets. Articles that compared diets with AGP (AGP+) or without AGP (AGP–) for pigs were collected from electronic databases and the performance results were entered in a database. A meta-analysis was performed following the sequence: graphical analysis, correlation, and variance-covariance. The performance results observed in the meta-analysis, feed cost, and AGP costs were used to build equations to estimate the economic effect of withdrawing AGP. The database comprised 81 scientific articles containing 103 experiments totalizing 42,923 pigs. Avilamycin (24.7 %) was the most frequent AGP in the database, followed by Colistin (15.4 %), Tiamulin (11.7 %), Tylosin (8.0 %), Lincomycin (9.4 %), and Bacitracin (5.4 %). Weight gain (p < 0.05) increased in AGP+ diets during post-weaning (6.5 %). However, there was no effect of AGP on weight gain of growing-finishing pigs. There was better (p < 0.05) feed conversion in pigs fed AGP+ diets in all rearing phases. Weight gain and feed conversion improved (p < 0.05) with the addition of Avilamycin, Bacitracin, and Tylosin. AGP withdrawal in the post-weaning phase increased feed costs by US$ 0.86 per animal and in growing-finishing phase the increase was US$ 3.11. Thus, pigs fed AGP+ diets have a better performance than pigs fed AGP- diets and the withdrawal of AGP increases feed costs.

Effect of Probiotics and Multi-Component Feed Additives on Microbiota, Gut Barrier and Immune Responses in Broiler Chickens During Subclinical Necrotic Enteritis.

The withdrawal of antibiotic growth promoters from poultry feed has increased the risk of necrotic enteritis (NE) outbreaks. This study examined the effects of a probiotic (PROB) or probiotic/prebiotic/essential oil supplement (PPEO) during a subclinical NE challenge. On day (d) of hatch, 960 male broilers were randomized to four groups (8 pens/treatment, 30 birds/pen) including (1) negative control (NC): corn-soybean meal diet; (2) positive control (PC): NC + 20 g Virginiamycin/ton diet; (3) NC + 227 g PROB/ton diet; and (4) NC + 453 g PPEO/ton diet. One d after placement, birds were challenged by a coccidia vaccine to induce NE. Feed intake and body weights were measured on d 8 (NE onset) and end of each feeding period. On d 8, the small intestines of three birds/pen were examined for NE lesions. Jejunum samples and ileal mucosal scrapings from one bird/pen were respectively collected to measure mRNA abundance (d 8 and d 14) and profile the microbiota (d 8 and d 42). Data were analyzed in JMP or QIIME 2 and significance between treatments identified by LSD (P < 0.05). PROB and PPEO had significantly lower mortality (d 0–14) and NE lesion scores compared to NC. Feed conversion ratio was significantly lower in PC, PROB, and PPEO, while average daily gain was higher in PPEO and PC groups compared to NC from d 0–42. On d 8 and d 14, mRNA abundance of claudin-3 was higher in PPEO compared to NC. On d 14, compared to NC, mRNA abundance of sIgA and PGC-1α in PROB and PPEO were lower and higher, respectively. Compared to NC, PPEO increased mTOR abundance on d 14. On d 8, relative abundance of Clostridium sensu stricto 1, Ruminiclostridium9, Prevotellaceae, Prevotellaceae UCG-014, ASF356, and Muribaculaceae was higher in NC compared to PPEO and PROB, while Lactobacillus was lower in NC. Escherichia-Shigella had higher abundance in PC compared to PPEO and PROB. Collectively, these data indicate that during a subclinical naturally occurring NE, supplementation of PROB or PPEO supports performance and reduces intestinal lesions, potentially through modifying tight junction proteins, gut microbiota, immune responses, and cell metabolism.

Effects of Eimeria maxima and Clostridium perfringens infections on cecal microbial composition and the possible correlation with body weight gain in broiler chickens

With the voluntary and regulatory withdrawal of antibiotic growth promoters from animal feed, coccidiosis and necrotic enteritis (NE) emerge as the top two enteric poultry infectious diseases responsible for major economic loss worldwide. The objective of this study was to investigate the correlation between the cecal microbiota compositions with the growth trait after coccidiosis and NE. In this study, the effects of Eimeria maxima and/or Clostridium perfringens infections on the microbial composition and potential correlation with the body weight gain were investigated in broiler chickens using 16S rRNA gene sequencing. E. maxima and C. perfringens coinfection successfully induced NE with its typical gut lesions and significant reductions in the percentage of relative body weight gain (RBWG%). The NE challenge model did not affect cecal microbial diversity, but influenced the cecal microbial composition. KEGG enzymes in microbiota were significantly altered in abundance following dual infections. Furthermore, significant correlations between cecal microbiota modules and RBWG% were identified in the sham control, E. maxima or C. perfringens infected groups. Understanding of host-microbiota interaction in NE would enhance the development of antibiotics-independent strategies to reduce the harmful effect of NE on the gut microbiota structure, and improve the gut health and poultry production.

Immunization with a subunit of housekeeping proteins provide partial protection against experimental necrotic enteritis in broiler chickens

Abstract Necrotic enteritis (NE) due to Clostridium perfringens (CP) infections is a prevalent enteric infectious disease accounting for $6 billion economical loss in global poultry industry. The increasing incidence of NE has been associated with the reduction and withdrawal of antibiotic growth promoters from animal feed during recent years. Therefore, the development of effective vaccines specific for NE assumes a priority for poultry industry. By far, no any effective vaccines are commercially available in broiler chickens. Our aim was to identify the potential CP proteins as vaccine targets for NE. We expressed 4 different recombinant CP proteins targeting 6 antigens: 3 major virulence-related toxins (alpha-toxin, NetB and TpeL), and 3 pilus subunit or enzymes (Collagen adhesion protein (Cna), Fructose-1,6-bisphosphate aldolase (FBA) and a Hypothetic protein (HP)), and their protection efficacies were evaluated with a severe Coccidiosis/NE challenge model using netB+tpeL+ CP strain. Young chicks were immunized twice subcutaneously with adjuvanted CP proteins on days 4, and 15. Prior to challenge, the chickens immunized with related antigens had much higher serum antibody titers for Cna and FBA/HP antigens than those with any of 3 toxin antigen-immunized groups. Following the challenge, the Cna-immunized group showed the least mortality and lowest lesion score among the individual antigen-vaccinated groups, while the pooled antigens-immunized group demonstrated no mortality against virulent challenges. The results indicate that the immunization with CP housekeeping protein, Cna, conferred better protection than other conventional virulence-related toxin proteins. Mucosal delivery system is being investigated next.

Comparison of the Pathogenicity of Five Clostridium perfringens Isolates Using an Eimeria maxima Coinfection Necrotic Enteritis Disease Model in Commercial Broiler Chickens

Clostridium perfringens (CP) is the etiologic agent of necrotic enteritis (NE) in broiler chickens that is responsible for massive economic losses in the poultry industry in response to voluntary reduction and withdrawal of antibiotic growth promoters. Large variations exist in the CP isolates in inducing intestinal NE lesions. However, limited information is available on CP isolate genetics in inducing NE with other predisposing factors. This study investigated the ability of five CP isolates from different sources to influence NE pathogenesis by using an Eimeria maxima (EM) coinfection NE model: Str.13 (from soil), LLY_N11 (healthy chicken intestine), SM101 (food poisoning), Del1 (netB+tpeL-) and LLY_Tpel17 (netB+tpeL+) for NE-afflicted chickens. The 2-wk-old broiler chickens were preinfected with EM (5 × 103 oocysts) followed by CP infection (around 1 × 109 colony-forming units per chicken). The group of the LLY_Tpel17 isolate with EM coinfection had 25% mortality. No mortality was observed in the groups infected with EM alone, all CP alone, or dual infections of EM/other CP isolates. In this model of EM/CP coinfections, the relative percentages of body weight gain showed statistically significant decreases in all EM/CP groups except the EM/SM101 group when compared with the sham control group. Evident gut lesions were only observed in the three groups of EM/LLY_N11, EM/Del1, and EM/LLY_Tpel17, all of which possessed an essential NE pathogenesis locus in their genomes. Our studies indicate that LLY_Tpel17 is highly pathogenic to induce severe gut lesions and would be a good CP challenge strain for studies investigating pathogenesis and evaluating the protection efficacy for antibiotic alternative approaches.

Galacto-Oligosaccharides Modulate the Juvenile Gut Microbiome and Innate Immunity To Improve Broiler Chicken Performance.

Improvements in growth performance and health are key goals in broiler chicken production. Inclusion of prebiotic galacto-oligosaccharides (GOS) in broiler feed enhanced the growth rate and feed conversion of chickens relative to those obtained with a calorie-matched control diet. Comparison of the cecal microbiota identified key differences in abundances of Lactobacillus spp. Increased levels of Lactobacillus johnsonii in GOS-fed juvenile birds at the expense of Lactobacillus crispatus were linked to improved performance (growth rate and market weight). Investigation of the innate immune responses highlighted increases of ileal and cecal interleukin-17A (IL-17A) gene expression counterposed to a decrease in IL-10. Quantification of the autochthonous Lactobacillus spp. revealed a correlation between bird performance and L. johnsonii abundance. Shifts in the cecal populations of key Lactobacillus spp. of juvenile birds primed intestinal innate immunity without harmful pathogen challenge.IMPORTANCE Improvements in the growth rate of broiler chickens can be achieved through dietary manipulation of the naturally occurring bacterial populations while mitigating the withdrawal of antibiotic growth promoters. Prebiotic galacto-oligosaccharides (GOS) are manufactured as a by-product of dairy cheese production and can be incorporated into the diets of juvenile chickens to improve their health and performance. This study investigated the key mechanisms behind this progression and pinpointed L. johnsonii as a key species that facilitates the enhancements in growth rate and gut health. The study identified the relationships between the GOS diet, L. johnsonii intestinal populations, and cytokine immune effectors to improve growth.

Withdrawal of antibiotic growth promoters from broiler diets: performance indexes and economic impact

This study aimed to estimate the productive and economic impacts caused by the withdrawal of antibiotic growth promoters (AGP) from broilers diet. Indexed publications that compared diets with or without AGP (AGP+/AGP−) for broilers (from initial to final phase) were collected and the results of feed intake, weight gain, and feed conversion were compiled in a database. A meta-analysis was performed following sequential analyses: graphical approach (to observe biological data coherence), correlation (to identify related factors), and variance-covariance (to compare groups). The annual number of broiler slaughtered in Brazil, target weight gain and feed conversion for each phase, the variation in feed conversion, feed cost, and AGP costs were used to build a model to estimate the effects of the AGP withdrawal on feeding costs. The database comprised 174 scientific articles containing 183 experiments, totaling 121,643 broilers, most of which were Ross (52% of the studies). The most frequent AGP sources/forms in the database were Avilamycin (41% of the AGP+ treatments), Flavomycin (19%), Virginiamycin (16%), and Bacitracin (14%). Higher feed intake, weight gain, and lower feed conversion were attributed (P < 0.05) to AGP+ diets during Initial phase (1 to 21 D). In Final phase (22 to 42 D) no differences were observed in performance variables. Treatments AGP+ presented higher weight gain and better feed conversion in the Total period (1 to 42 D). The results of feed conversion were improved (P < 0.05) with Avilamycin and Flavomycin; Virginiamycin improved weight gain and feed conversion. In the Total period, the economic impact was $0.03 per animal and a total of $183,560,232 per year. It was concluded that broilers fed AGP+ diets have higher weight gain and better feed conversion than those fed AGP− diets, and withdrawing AGP increases production costs.

Characterization of Clostridium perfringens Strains Isolated from Healthy and Necrotic Enteritis-Afflicted Broiler Chickens.

Necrotic enteritis (NE) is an important enteric disease in poultry, and Clostridium perfringens (CP) type A strains are the primary etiology. NE is responsible for annual losses of US $6 billion to the poultry industry in the United States. An increase in the incidence of NE has been also associated with withdrawal of antibiotic growth promoters from poultry feed. In this study, CP strains isolated from healthy and NE-afflicted birds were characterized microbiologically and molecularly, and their virulence was experimentally tested in chickens. All strains were hemolytic, lecithinase positive, and identified as CP by biochemical tests. Three distinct colony morphologies were seen in brain-heart infusion media with 0.3% agarose, FeSO4, and ZnCl2. The CP strains responded differently to iron chelation with 2,2'-bidypinol. PCR toxinotyping showed that all tested strains were alpha toxin-positive, seven (N11, N10, CP1, CP5, CP13, JGS, and Del1) were beta2-toxin-positive, and only one (Del1) was necrotic enteritis toxin B-like-positive. In vivo studies indicated that most isolates, including strain N11 isolated from the normal chicken gut, were sufficiently virulent to produce NE disease in the Eimeria/CP dual infection model. The Del1 and N11 strains merit further investigation to identify their virulence factors and immune-protective antigens.

PROBIOTYKI, PREBIOTYKI I SYNBIOTYKI - SPOS�B DZIALANIA, OGRANICZENIA I OSIAGNIECIA

The withdrawal of antibiotic growth promoters from poultry industry have forced farmers to seek alternatives for the posing a risk factors of cross-resistance acquisition by harmful bacteria. A particular nuisance became salmonellosis and campylobacteriosis forcing to the elimination of whole poultry flocks as well as causing dangerous zoonotic diseases in humans. An excellent replacement for antibiotics have become the pro-, pre-and synbiotic substances which have a beneficial effect on the host organism through the development intensification of healthy intestinal microbial strains and the elimination of pathogenic strains. Such preparations may be administered both in the water spray as well as in feed. Excellent and promising method appears to be their injection directly into the egg air chamber in the 12th day of incubation. However, further studies are required to determine the appropriate doses as well as combinations of bioactive substances and to determine the optimal way for their delivery.