The Beneficial Effects of Components of Garlic (Allium sativum L.) in the Poultry Industry

Plant-based antimicrobial agents are readily available, cost-effective, and exhibit low toxicity, making them promising alternatives in combatting microbial infections. Among these plants, garlic (Allium sativum) stands out for its traditional medicinal use in effectively combating various microorganisms. In the poultry industry, preventing avian virus and bacterial infections is paramount for chicken husbandry. However, using conventional drugs poses potential risks to human health. Garlic, a widely used Asian plant in traditional medicine for various pathologies, has shown potential as an herbal prophylactic remedy against viral and bacterial infections. Recently, researchers explored garlic and its derivatives as a scientific strategy in veterinary practices for diverse purposes, such as improving poultry production characteristics and acting as antibiotic growth promoters. This comprehensive review delves into garlic and its derivatives as preventive and corrective treatments for viral diseases in laying hens and broilers. The paper highlights their potential effectiveness and safety as a natural means to enhance poultry health and welfare while mitigating the risks associated with conventional drug usage in the food industry.

Effects of methionine supplementation in a reduced protein diet on growth performance, oxidative status, intestinal health, oocyst shedding, and methionine and folate metabolism in broilers under Eimeria challenge

Background This study investigated effects of different methionine(Met)supplementation levels in a reduced protein diet on growth performance,intestinal health,and different physiological parameters in broilers under Eimeria challenge.A total of 600 fourteen-day-old Cobb500 male broilers were challenged with E.maxima,E.tenella,and E.acervulina,and randomly allocated in a 2×5 factorial arrangement.Birds received normal protein diets(20%crude protein,NCP)or reduced protein diets(17%crude protein,LCP),containing 2.8,4.4,6.0,7.6,and 9.2 g/kg of Met.Results On 6 and 9 days post inoculation(DPI),increasing Met level linearly improved the growth performance(P<0.05).Total oocyst shedding linearly increased as Met level increased(P<0.05).Duodenal villus height(VH):crypt depth(CD)in the LCP groups were higher on 6 DPI(P<0.01)while lower on 9 DPI(P<0.05)compared to the NCP groups.Jejunal CD and duodenal VH:CD changed quadratically as Met level increased(P<0.05).On 6 DPI,liver glutathione(GSH)and glutathione disulfide(GSSG)linearly increased as Met level increased(P<0.05).On 9 DPI,GSSG quadratically increased,whereas GSH:GSSG quadratically decreased as Met levels increased(P<0.05).The expression of amino acid transporters linearly decreased as Met level increased(P<0.05).The expression of zonula occludens 2 and claudin-1 linearly increased on 6 DPI whereas decreased on 9 DPI as Met level increased(P<0.05).The expressions of cytokines were lower in the LCP groups than the NCP groups(P<0.05).Interaction effects were found for the expression of IL-10 and TNFαon 6 DPI(P<0.05),where it only changed quadratically in the NCP group as Met level increased.The expression of Met and folate metabolism genes were lower in the LCP groups than the NCP groups on 9 DPI(P<0.05).The expression of these genes linearly or quadratically decreased as Met level increased(P<0.05).Conclusion These results revealed the regulatory roles of Met in different physiological parameters including oxidative status,intestinal health,and nutrient metabolism in birds fed reduced protein diet and challenged with Eimeria.

Increased CO2 Levels during the First Half of Incubation at High Altitude Modifies Embryonic Development of Fertile Leghorn Breeder Eggs

The exchange of oxygen (O2) and carbon dioxide (CO2) within an incubator has a significant impact on embryonic development (ED) and hatching processes. This study examines the influence of non-ventilation (NV) conditions during the first ten days of incubation at high altitudes on Leghorn hens hatching eggs. Five hundred four hatching eggs were equally divided into three treatment groups and placed in twelve incubators (R = 4). The first group was subjected to standard ventilated conditions (V) during the setting phase. The ventilation inlet holes of the remaining incubators in the NV treatments were closed with either micropore (M) or polypropylene (P) tape, referred to as NVM and NVP groups, respectively. These two different airtight settings were intended to allow for a gradual rise in CO2 naturally generated by the embryos. Results indicate that carbon dioxide concentration gradually increased during the first half of incubation, reaching 1.42% in the NVM group and 1.20% in the NVP group, while the V condition group remained at 0.15%. From 10 days of incubation onwards, normal V conditions were restored in all incubators. The highest hatchability of fertile eggs (HFE) was shown by the NVP group (55.7%), followed by the V (52.6%) and NVM (38.6%) groups. The NVP group showed a greater yolk-free body mass (YFBM) from 10 days of incubation until the hatch basket transfer. NV conditions during the first 10 days of incubation at high altitude produced higher YFBM with gradually decreasing yolk sac mass. In comparison to the NVM and V conditions, the particular NVP condition showed a beneficial impact on the quality of hatched chicks. Sustaining NVP condition (1.2% of CO2) throughout the first half of incubation at high altitude generated the optimal environment in the incubator ensuring the best hatchability results. This study highlights how important it is for hatchery managers to recognize the influence of low O2 and high levels of CO2 on the development trajectories of Leghorn embryos during early incubation at high altitudes.

Correlation between Intestinal Health and Coccidiosis Prevalence in Broilers during Different Seasons of the Year in Brazil from 2012 to 2018

Coccidiosis is a disease caused by intracellular protozoan parasites, specifically belonging to the genus Eimeria. These parasites target the gastrointestinal tract in different types of hosts, causing sores in the intestinal lining. The presence of these lesions reduces the animal’s ability to digest and absorb nutrients, significantly impacting their overall performance. The current study aimed to explore the potential correlation between seasonal variations and the incidence of Eimeria spp-induced lesions in broiler chickens’ gastrointestinal tracts in Brazil from 2012 to 2018. A total of 8,607 broiler chickens, aged 14 to 42 days, were sampled from 103 poultry integrated companies in Brazil to conduct intestinal health examinations. The sampling process involved selecting 3 to 6 chickens from each poultry house for examination. The assessment included various abnormalities such as shedding of intestinal cells and excessive fluid and mucus presence, thickening and tension of the intestines, food movement, roughened mucosal surface resembling a Turkish towel in the small intestine, tissue death, duodenal inflammation, intestinal inflammation, gizzard erosion, presence of worms and bedding material, and ingestion of mealworms. During the seasons, winter exhibited the highest average occurrence of Eimeria maxima microorganisms at 52.83%, with E. acervulina following closely at 26.42% in second place. In spring, E. maxima had an occurrence of 11.31%, while in fall, E. tenella had the lowest occurrence at 6.74%. When analyzing the seasonal occurrence of Eimeria, it was observed that E. maxima micro was more common during winter compared to summer (P = 0.0491). However, no discernible variation was observed in the occurrence of the remaining species across different seasons. Research findings suggest that subclinical coccidiosis is most prevalent during the winter season in Brazil. Likewise, clinical disease caused by E. acervulina is also prevalent during this time. In contrast, E. maxima is more likely to cause clinical disease in the spring, whereas E. tenella is more commonly associated with clinical disease in the fall. Lesions induced by Eimeria spp. are associated with factors influencing the overall health of broiler intestines. These findings allow for the utilization of seasonal metrics in disease management, thereby reducing economic losses associated with the condition.

Supranutrition of microalgal docosahexaenoic acid and calcidiol improved growth performance,tissue lipid profiles,and tibia characteristics of broiler chickens

Background Docosahexaenoic acid(DHA)and calcidiol could be enriched in chicken for improving public nutrition and health.It remains unclear if supranutritional levels of DHA and calcidiol impair growth performance or metabolism of broiler chickens.This study was to determine singular and combined effects of high levels of supplemental DHA-rich microalgal biomass or oil and calcidiol on growth performance,concentrations of triglycerides,cholesterol,and nonesterfied fatty acids in plasma,liver,breast,and thigh,and biophysical properties of tibia.Methods In Exp.1,144 day-old Cornish chicks were divided into 4 groups(6 cages/treatment,6 birds/cage),and were fed a corn-soybean meal basal diet(BD),BD+10,000 IU calcidiol/kg(BD+Cal),BD HA),and BD+1%DHA-rich Aurantiochytrium(1.2 g DHA/kg;BD+D+Cal+DHA for 6 weeks.In Exp.2,180 day-old chicks were divided into5 groups,and were fed:BD,BD+DHA(0.33%to 0.66%oil,1.5 to 3.0 g DHA/kg),BD EPA/kg),BD+DHA+EPA(1.9%to 3.8%eicosapentaenoic acid-rich Nannochloropsis sp.CO18,0.3 to 0.6 g BD+DHA+calcidiol(6000 to 12,000 IU/kg diet),and+DHA+EPA+Cal for 6 weeks.Results Birds fed BD+Cal diet in Exp.1 and BD lower(P+DHA<0.05)+EPA diet in Exp.2 had higher(P<0.05)body weight gain(10%-11%)and gain:feed ratio(7%),and total cholesterol and triglyceride concentrations in plasma(18%-54%),liver(8%-26%),breast(19%-26%),and thigh(10%-19%),respectively,over the controls.The two diets also improved(P<0.05)tibial breaking strength(8%-24%),total bone volume(2%-13%),and(or)bone mineral density(3%-19%)of chickens.Conclusion Supranutrition of dietary calcidiol and DHA alone or together did not produce adverse effects,but led to moderate improvements of growth performance,lipid profiles of plasma and muscle,and bone properties of broiler chickens.

Comparison of Chick Quality, Health, and Inflammation from Two Hatchery Environments

Hatchery contamination can result in ingested or inhaled microbes that may modify colonization of the intestinal and respiratory tract, with potential to influence early growth, inflammation, and overall health. Six experiments were completed to compare chick quality, inflammation, and health between two hatcheries (H1 and H2). On embryonic d0, 45 eggs from the same breeder flock were set at each hatchery. On d0, length, abdominal height, navel and leg abnormalities, and self-righting were measured for 36 chicks/hatchery, yolk sacs were weighed, and crop/cloaca swabs were cultured from 12 chicks/hatchery. On d7, mid-ileum and ceca were cultured from 12 chicks/hatchery. On d0 and d7, body weight (BW) and intestinal weight were measured, lung/air sac swabs and liver were cultured, and liver and air sacs were scored for health. Blood was collected on d0 and d7 for serum alpha-1-acid glycoprotein concentrations in Exp 1 and 2. Data was analyzed using Student’s t-test or χ2, significance p < 0.05. On d0, BW, length, yolk free BW, and intestinal weight were greater for H2 chicks (p < 0.05). Liver bacterial recovery was decreased in H2 on d0 (p < 0.05) and there were fewer average leg and righting abnormalities in H2 (p < 0.05). Decreased lactase positive Enterobacteriaceae were noted in H2 in crop/cloaca and lung/air sac swabs (p < 0.05), and of alpha and beta hemolysis in crop/cloaca swabs, and alpha and gamma hemolysis in lung/air sac swabs (p < 0.05) on d0. By d7, only alpha hemolytic bacteria were increased in lung/air sac swabs of H2. Based on factors measured, chicks from H2 showed favorable microbial colonization, starting quality, and improved health on d0. While not sustained through d7, differences in d0 microbial recovery may have shifted microbial development and potentially influenced immune response development. These experiments elucidated the importance of hatchery environment on early chick quality, microbial colonization, overall inflammation, and chick health.

Essential Oils as an Alternative to Antibiotics to Reduce the Incidence and Severity of Necrotic Enteritis in Broiler Chickens: A Short Review

Due to the removal of antibiotic growth promoters (AGPs) and consumer pressure for antibiotic-free (ABF) or no antibiotics ever (NAE) poultry production, there is a need for sustainable alternatives to prevent disease in commercial poultry operations. Without AGPs, there has been a rise in diseases that were traditionally controlled by subtherapeutic levels of antibiotics in the diet. This has impacted the health of commercial poultry and has been a significant cost to poultry producers. To mitigate this, the industry has started to investigate alternatives to antibiotics to treat these forthcoming health issues, such as necrotic enteritis (NE). NE is an enteric disease caused by an over proliferation of toxigenic Clostridium perfringens (CP) in the gastrointestinal tract. Although CP is a commensal in the avian intestinal tract, dysbiosis caused by inflammation and impaired intestinal integrity facilitates uncontrolled replication of CP. Infectious agents, such as Eimeria maxima, appear to be a predominant predisposing factor that promotes NE. However, non-infectious stressors, including dietary changes, have also been associated with NE to some degree. As a result of increased pressure to restrict the use of antibiotics, there is a need for research evaluating the efficacy of alternatives, such as plant-derived essential oils, as potential tools to mitigate NE in commercial poultry flocks. The aim of this study is to review the effects of essential oils as an alternative to antibiotics to reduce the incidence and severity of necrotic enteritis in broiler chickens.

Isolation and Progeny Transmission of Non-Temperature-Sensitive MS-H Vaccine Strains of Mycoplasma synoviae from Temperature-Sensitive MS-H-Vaccinated Laying Breeder Hens

This pilot study reports the vertical transmission and reverse thermosensitivity of the MS-H vaccine strain of Mycoplasma synoviae (MS) by RAPD in commercial breeders and their progeny. At two weeks of age, breeders were vaccinated with the ts+ MS-H strain. At 9 weeks of age, an outbreak of infectious synovitis (IS) was detected in the progeny. Tracheal swab samples were collected from breeders at 24, 39, 48, and 70 weeks of age. At 9 weeks, pullets swab from the elbow joints were collected. RAPD was performed on the isolates at 39.5°C, and the same ts- MS-H strains were identified in the breeder hens and their progeny. Tracheal swabs from breeder hens were negative to MS isolation at 37°C and 39.5°C at 24- and 39-weeks. MS isolation was recovered from tracheal swabs from 9/10 and 10/10 breeders at 48- and 70- week. At 9 weeks of age in the progeny, MS was isolated from tracheal swabs of 10/10 from non-IS pullets. MS was isolated from 9/10 joints samples. The isolates from breeder hens and their progeny showed non-significant differences in five antimycoplasmic MIC100 values;otherwise, enrofloxacin presented a significant difference in MIC100 value (p < 0.05). This investigation demonstrated the reversal of the thermosensitivity, pathogenicity, and vertical transmission of the MS-H strain. Consequently, it is crucial to contemplate the danger of reversing pathogenicity and transmission to progeny when applying the MS-H vaccine strain.

具有可生物降解、细胞相容、抗菌及生物膜控制的壳聚糖磺基甜菜碱衍生物薄膜生物材料的研究

The purpose of this research was to develop a chitosan sulfobetaine(CS-SNCC)film via the solutioncasting method as a biodegradable antibacterial material for biomedical applications.Chitosan and monochloro-triazine sulfobetaine were used as the raw materials for CS-SNCC preparation,and Fourier-transform infrared(FTIR),ultraviolet–visible(UV-Vis),energy-dispersive X-ray(EDX),and Xray photoelectron spectroscopy(XPS)spectra were used to characterize and analyze the structure of the synthesized CS-SNCC.Furthermore,the swelling property,thermal stability,biodegradability,cytocompatibility,and antibacterial properties of the CS-SNCC film were comprehensively investigated and compared with those of the chitosan film.The results for the film’s enzymatic biodegradation behavior show that the CS-SNCC film undergoes a weight loss of 45.54%after 21 days of incubation.In addition,the CS-SNCC film effectively resists bacterial adhesion,prevents the formation of bacteria biofilms,and exhibits high antibacterial activity,with inactivation rates of 93.43%for Escherichia coli and 91.00%for Staphylococcus aureus.Moreover,the CS-SNCC film shows good cellular activity and cytocompatibility according to the cytotoxicity results.Therefore,the prepared biodegradable,cytocompatible,antibacterial,and biofilm-controlling CS-SNCC film has potential for biomedical applications.

Comprehensive analysis of coding and non-coding RNA transcriptomes related to hypoxic adaptation in Tibetan chickens

Background:Tibetan chickens,a unique native breed in the Qinghai-Tibet Plateau of China,possess a suite of adaptive features that enable them to tolerate the high-altitude hypoxic environment.Increasing evidence suggests that long non-coding RNAs(lncRNAs)and microRNAs(miRNAs)play roles in the hypoxic adaptation of high-altitude animals,although their exact involvement remains unclear.Results:This study aimed to elucidate the global landscape of mRNAs,lncRNAs,and miRNAs using transcriptome sequencing to construct a regulatory network of competing endogenous RNAs(ceRNAs)and thus provide insights into the hypoxic adaptation of Tibetan chicken embryos.In total,354 differentially expressed genes(DE genes),389 differentially expressed lncRNAs(DE lncRNAs),and 73 differentially expressed miRNAs(DE miRNAs)were identified between Tibetan chickens(TC)and control Chahua chickens(CH).GO and KEGG enrichment analysis revealed that several important DE miRNAs and their target DE lncRNAs and DE genes are involved in angiogenesis(including blood vessel development and blood circulation)and energy metabolism(including glucose,carbohydrate,and lipid metabolism).The ceRNA network was then constructed with the predicted DE gene-DE miRNA-DE lncRNA interactions,which further revealed the regulatory roles of these differentially expressed RNAs during hypoxic adaptation of Tibetan chickens.Conclusions:Analysis of transcriptomic data revealed several key candidate ceRNAs that may play high-priority roles in the hypoxic adaptation of Tibetan chickens by regulating angiogenesis and energy metabolism.These results provide insights into the molecular mechanisms of hypoxic adaptation regulatory networks from the perspective of coding and non-coding RNAs.

Isolation, screening and identifi cation of Bacillus spp. as direct-fed microbial candidates for aflatoxin B_1 biodegradation

Objective: To evaluate the ability of Bacillus spp. as direct-fed microbials(DFM) to biodegrade al atoxin B1(AFB1) by using an in vitro digestive model simulating in vivo conditions.Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 μg/m L of AFB1 in modii ed Czapek-Dox medium. Plates were incubated at 37 °C and observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16 S r RNA sequence analyzes for identii cation. Tolerance to acidic p H, osmotic concentrations of Na Cl, bile salts were tested, and antimicrobial sensitivity proi les were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time(3:15 h), supernatants and digesta were collected for high-performance liquid chromatography l uorescence detection analysis by triplicate.Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of l uorescence and area of clearance around each colony in modii ed Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identii ed the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions(p H 2.0), tolerant to a high osmotic pressure(Na Cl at 6.5%), and were able to tolerate 0.037% bile salts after 24 h of incubation. No signii cant dif erences(P > 0.05) were observed in the concentrations of AFB1 in neither the supernatants nor digesta samples evaluated by highperformance liquid chromatography with l uorescence detection between positive control or DFM treated groups. Conclusions: In vitro digestion time was not enough to confirm biodegradation of AFB1. Further studies to evaluate the possible biodegradation ef ects of the BacillusDFM when continuously administered in experimentally contaminated feed with AFB1, are in progress.

Supplementation of amylase combined with glucoamylase or protease changes intestinal microbiota diversity and benefits for broilers fed a diet of newly harvested corn

Background: The effect of amylases combined with exogenous carbohydrase and protease in a newly harvested corn diet on starch digestibility, intestine health and cecal microbiota was investigated in broiler chickens.Methods: Two hunderd and eighty-eight 5-day-old female chickens were randomly divided into six treatments: a newly harvested corn-soybean meal diet(control); control supplemented with 1,500 U/g α-amylase(Enzyme A);Enzyme A + 300 U/g amylopectase + 20,000 U/g glucoamylase(Enzyme B); Enzyme B + protease 10,000 U/g(Enzyme C); Enzyme C + xylanase 15,000 U/g(Enzyme D); and Enzyme D + cellulase 200 U/g + pectinase 1,000 U/g(Enzyme E). Growth performance, starch digestibility, digestive organ morphology, and intestinal microbiota were evaluated in the birds at 16 and 23 d of age.Results: Compared with the control diet, supplementation with Enzyme A significantly decreased ileum lesion scoring at 16 d of age(P < 0.05); supplementation with Enzyme B or Enzyme C showed positive effects on ileal amylopectin and total starch digestibility(P < 0.05); Broilers fed with a diet supplemented with Enzyme D had a tendency to decrease body weight gain at 23 d. Enzyme E supplementation improved lesion scoring of jejunum and ileum at 16 d(P < 0.05), and increased ileal amylopectin or total starch digestibility at 23 d(P < 0.05).Supplementation of enzymes changed cecal microbiota diversity. High numbers of Campylobacter, Helicobacter and Butyricicoccus, Anaerostipes and Bifidobacterium, Sutterella and Odoribacter were the main genera detected in supplementations with Enzymes B, C, D, and E respectively.Conclusions: Supplementation with amylase combined with glucoamylase or protease showed a beneficial effect on starch digestibility and intestinal microbiota diversity, and increased growth of broilers fed with newly harvested corn.

Dried fruit pomace inclusion in poultry diet:growth performance, intestinal morphology and physiology

Background: Fruit pomaces are by-products rich in polyphenol compounds and dietary fiber. They seem to play an important role in regulating the gut microbiota, morphology and physiology. The aim of this study was to assess whether apple(A), blackurrant(B) or strawberry(S) pomaces could be suitable ingredients in broiler diets and their effect on gut health. A total of 480 male broilers were randomly allotted to 8 dietary treatments with lower(3%-L) or higher(6%-H) dietary fiber content: two control groups(CL/CH), two A diets(AL/AH), two B diets(BL/BH), two S diets(SL/SH). Diet and fruit pomaces were chemically analyzed to assess polyphenol concentration and fibre fraction content. After the evaluation of growth performance, 6 birds/group were slaughtered at 35 days of age. Morphometric and histopathological investigations were performed on duodenum, jejunum and ileum.Excreta were collected to perform microbiota evaluation by 16 S DNA sequencing. Weight, viscosity, enzymatic activity, short chain fatty acid(SCFAs) and ammonia concentration were determined in ileum and/or ceca content.Results: A pomace and A diets showed the lowest polyphenol content and the highest content of soluble fibre fraction. No significant differences were observed for growth performance, gut morphometry and histopathology(P > 0.05). Dietary fruit pomace inclusion increased the weight of ileum and ceca and the ileum digesta viscosity(P < 0.05). In the ileum, A and S groups showed lower bacterial α-glucosidase activity than C groups. Moreover,small intestine SCFAs concentration was higher in fruit pomaces diets(P < 0.05). In ceca, B and S groups showed lower ammonia concentration and higher SCFAs than C. Dietary treatments also influenced the activity of α-glucosidase, α-galactosidase, β-galactosidase β-glucuronidase and xylase. Regarding microbiota, at phylum level,Firmicutes were differentially abundant across treatment(maximum for C and minimum in S, FDR > 0.05). At genus level, an increase of Weissella in AH and Erwinia in S/B diets, as well as a decrease of Lactobacillus in all fruit pomace groups were recorded(P < 0.05).Conclusions: Fruit pomaces could be suitable ingredients in poultry nutrition even if further studies are needed to better understand which doses is more recommended to avoid negative effects on gut microbiota.

Effects of 50 Percent Substitution of Soybean Meal by Alternative Proteins from <i>Hermetia illucens</i>or Spirulina platensis in Meat-Type Chicken Diets with Graded Amino Acid Supply

Alternative protein sources, such as insects or algae meals are in special focus of animal nutrition in order to replace soybean meal (SBM). As part of the multidisciplinary project “sustainability transitions” this study evaluated effects of replacing SBM by partly defatted larvae meal from the black soldier fly, Hermetia illucens or meal from the micro algae Spirulina platensis in broiler diets. The aim of the current study was to investigate the chickens’ growth performance and the intestinal morphology as well as the health status. 288 one-day-old male growing chickens (Ross 308) from a commercial hatchery were randomly allotted to 48 pens (6 birds per pen) for the growth study with five diets and feed supply on free choice level. The control diet was based on wheat, corn and SBM. The experimental diets replaced 50% of SBM by the alternative proteins under study, both on a basic level of amino acid (AA) supplementation (Lys and Met added equal to the control diet) (diet HM and SM) and an extended level of AA fortification (Lys, Met, Thr, Arg, Val added) (diet HM+ and SM+). Response of chickens was evaluated by zoo-technical parameters, which were under weekly control (e.g. growth, feed intake, feed conversion ratio, mortality). After finishing the growth study birds from control and the experimental diets at the basic level of AA supplementation were slaughtered after 12 hour fastening and utilized for gut morphometric analysis and histological evaluation of the health status. Diets at a basic level of AA fortification led to significant depression of growth, feed intake, feed conversion ratio, and protein conversion ratio, respectively. In addition, the acceptance of the Spirulina diet was lower (p ≤ 0.001) as compared to the Hermetia based diet. However, the extended level of AA supplementation improved all parameters significantly. Diets without extended AA supply altered some morphological parameters of the intestinal wall, but the nutritional significance of this observation needs to be verified in AA balanced diets. The health state of chicken was not impaired by the diets under study.

Evaluation of curcumin and copper acetate against Salmonella Typhimurium infection,intestinal permeability,and cecal microbiota composition in broiler chickens

Background:Interest in the use of natural feed additives as an alternative to antimicrobials in the poultry industry has increased in recent years because of the risk of bacterial resistance.One of the most studied groups are polyphenolic compounds,given their advantages over other types of additives and their easy potentiation of effects when complexes are formed with metal ions.Therefore,the objective of the present study was to evaluate the impact of dietary supplementation of copper acetate(CA),curcumin(CR),and their combination(CA-CR)against Salmonella Typhimurium colonization,intestinal permeability,and cecal microbiota composition in broiler chickens through a laboratory Salmonella infection model.S.Typhimurium recovery was determined on day 10 post-challenge by isolating Salmonella in homogenates of the right cecal tonsil(12 chickens per group)on Xylose Lysine Tergitol-4(XLT-4)with novobiocin and nalidixic acid.Intestinal integrity was indirectly determined by the fluorometric measurement of fluorescein isothiocyanate dextran(FITC-d)in serum samples from blood obtained on d 10 post-S.Typhimurium challenge.Finally,microbiota analysis was performed using the content of the left caecal tonsil of 5 chickens per group by sequencing V4 region of 16S rRNA gene.Results:The results showed that in two independent studies,all experimental treatments were able to significantly reduce the S.Typhimurium colonization in cecal tonsils(CT,P<0.0001)compared to the positive control(PC)group.However,only CA-CR was the most effective treatment in reducing S.Typhimurium counts in both independent studies.Furthermore,the serum fluorescein isothiocyanate dextran(FITC-d)concentration in chickens treated with CR was significantly lower when compared to PC(P=0.0084),which is related to a decrease in intestinal permeability and therefore intestinal integrity.The effect of dietary treatments in reducing Salmonella was further supported by the analysis of 16S rRNA gene sequences using Linear discriminant analysis effect size(LEfSe)since Salmonella was significantly enriched in PC group(LDA score>2.0 and P<0.05)compared to other groups.In addition,Coprobacillus,Eubacterium,and Clostridium were significantly higher in the PC group compared to other treatment groups.On the contrary,Fecalibacterium and Enterococcus in CR,unknown genus of Erysipelotrichaceae at CA-CR,and unknown genus of Lachnospiraceae at CA were significantly more abundant respectively.Conclusions:CR treatment was the most effective treatment to reduce S.Typhimurium intestinal colonization and maintain better intestinal homeostasis which might be achieved through modulation of cecal microbiota.

Deletions in the pyruvate pathway of Salmonella Typhimurium alter SPI1-mediated gene expression and infectivity

Background： Salmonella enter/ca serovar Typhimurium is a major foodborne pathogen worldwide. S. Typhimurium encodes type III secretion systems via Salmonella pathogenicity islands （SPI）, producing the major effector proteins of virulence. Previously, we identified two genes of Salmonella pyruvate metabolism that were up-regulated during chicken cell infection： pyruvate formate lyase I （pf/B） and b/functional acetaldehyde-CoA/alcohol dehydrogenase （adhE）. We were therefore interested in examining the role these genes may play in the transmission of Salmonella to humans. Methods： Mutant strains of Salmonella with single gene deletions for pflB and adhE were created. Invasion and growth in human HCT-8 intestinal epithelial cells and THP-1 macrophages was examined. Quantitative PCR was performed on 19 SPI-1 genes. Results： In HCT-8 cells, both mutant strains had significantly higher intracellular counts than the wild-type from 4 to 48 h post-infection. Various SPI-1 genes in the mutants were up-regulated over the wild-type as early as 1 h and lasting until 24 h post-infection. In THP-1 cells, no significant difference in internal Salmonella counts was observed; however, SPI-1 genes were largely down-regulated in the mutants during the time-course of infection. We also found five SPI-1 genes - hilA, hiIC hill）, sicP and rtsA - which were up-regulated in at least one of the mutant strains in log-phase broth cultures alone. We have therefore identified a set of SPI-1 virulence genes whose regulation is effected by the central metabolism of Salmonella.

The Effect of a<i>Lactobacillus</i>-Based Probiotic for the Control of Necrotic Enteritis in Broilers

Necrotic Enteritis (NE) caused by Clostridium perfringens (CP) in poultry is probably the most important bacterial disease in terms of economic implications. The disease is multi-factorial and is invariably associated with predisposing factors. The present study investigated the effect of a commercially available Lactobacillus-based probiotic (FM-B11) for the control of necrotic enteritis in broiler chickens. In experiment 1, one-day-of-hatch broiler chicks were randomly allocated to the following treatment groups: 1) Non-challenged (NC);2) Challenged (C);3) Challenged + probiotic (C+ FM-B11). Prior to placement, chicks in groups 2 and 3 received 0.25 mL of Salmonella typhimurium (ST) containing 105 cfu of viable cells by oral gavage. At 14, 15 and 16 days of age, all chicks in group 3 were treated with FM-B11 in the drinking water at a concentration of 106 cfu/ml. At 21d of age, all chicks in groups 2 and 3, were individually challenged with 5 × 104 sporulated oocysts of E. maxima by oral gavage. At 26d of age, all chicks in groups 2 and 3, were individually challenged with 108 cfu CP;body weight (BW) was recorded prior to challenge. The experiment was terminated at 29 days of age and the following parameters were evaluated: NE-associated mortality, CP lesion scores, CP concentrations in ileum, BW, and body weight gain (BWG). Chicks treated with FM-B11 had significantly (P < 0.05) higher body weight gain after challenge when compared to control challenge chickens. Total mortality was higher in the C group (48.8%) when compared to the C + FM-B11 (12.7%). Even though there was no significant (P > 0.05) difference in lesion score between C and C + FM-B11, group C + FM-B11 had significantly (P < 0.05) lower total number of cfu of CP recovered from the ileal mucosa and content samples when compared to group C. Experiment 2 was a unique and remarkable case report of a field outbreak of NE in a commercial broiler farm in Argentina. A reduction and control of the mortality associated with NE following 3 days of administration of FM-B11 was observed as compared with the control non treated house. These results imply that the commercially available Lactobacillus-based probiotic FM-B11 was able to reduce the severities of NE, as a secondary bacterial infection, in an experimental NE challenge model;as well as, in a commercial field outbreak of NE.

Supplementation of Vitamin E and C Reduces Oxidative Stress in Broiler Breeder Hens during Summer

The present experiment was carried out to evaluate the effects of vitamin E and C supplementation on the antioxidant status of broiler breeder hens during summer. Two hundred seventy broiler breeder birds (coloured) were divided into nine groups with three replicates having 10 birds in each, and were supplemented two levels of antioxidant vitamins E (250 mg or 500 mg/kg) or C (200 mg or 400 mg/kg) alone or in combinations for a period of 8 weeks. All the vitamin supplemented groups showed significantly lowered malondialdehyde (MDA) level, higher activities of superoxide dismutase (SOD), catalase (CAT) enzymes and higher Ferric Reducing Antioxidant Power (FRAP) activities in erythrocytes than control. However, supplementation with combination of vitamins proved to be better than individual supplementation. It could be concluded that the combination of vitamins C and E at lower level (@ 200 mg/kg and 250 mg/kg, respectively) had higher erythrocytic antioxidant status in coloured broiler breeder hens as compared to individual vitamin supplementation during hot and humid stress.

Comparison of total immunoglobulin A levels in different samples in Leghorn and broiler chickens

Objective: To standardize an ELISA protocol to quantify total immunoglobulin A(IgA)from different biological samples.Methods: Two independent experiments were conducted. In Experiment 1, total IgA levels were quantified from the lachrymal fluid, tracheal swab, and cloacal swab at various time points from Days 30 to 89 in white Leghorn chickens. Experiment 2 was conducted to evaluate the effect of 50 or 500 ppb of aflatoxin B1(AFB1) on total IgA quantified in samples from the lachrymal fluid, tracheal swab, gut content and cloacal swab in broiler chickens at 21 days of age.Results: Lachrymal fluid contained the highest level of IgA; however, the sampling procedure was time consuming and stressful to the bird, and the sample volume depends largely on the size of the chicken. Cloacal swabs also contained a high concentration of IgA; this sampling procedure was faster than lachrymal fluid sampling and was not affected by the age of the bird. Tracheal sampling was more difficult than cloacal sampling; the age of the bird limited the sampling, and the IgA concentration was the lowest detected at all sampling ages. 500 ppb of AFB1 significantly reduced total IgA concentration in the gut content compared with control or 50 ppb of AFB1 treated groups.Interestingly, a significant reduction in total IgA was also observed in those chickens that received 50 ppb of AFB1 in gut rinse when compared with cloacal swabs.Conclusions: The results of this study suggest that cloacal swab is an easy and reliable way to evaluate mucosal IgA concentration in both Leghorn and broiler chickens.

Insights into the proteomic profile of newly harvested corn and metagenomic analysis of the broiler intestinal microbiota

Background:The use of newly harvested corn in feed causes wet droppings in broilers and increased feed cost which was termed as“new season grain problem”.The present study was conducted to evaluate the proteomic profile of newly harvested corn and the subsequent influence on intestinal microbiol community for broiler chickens.Methods:Newly harvested corn stored for either half a month(HM)or two months(TM)was used,and the pasting properties,total soluble sugars,and proteomic analysis technology was used to explore the influence of storage on natural aging corn properties.Additionally,seventy-two 7-day-old Ross 308 male broiler chicken were fed with different stored corn.Apparent metabolizable energy(AME),digesta viscosity,intestinal morphology and microbiota were examined to explore the influence of feed corn storage on broiler chickens.Results:Pasting properties in the TM corn exhibited decreased viscoelastic properties.Proteomic studies found a total of 26 proteins that were differentially expressed between the two treatment groups.Proteins involved in starch and polysaccharides biosynthesis were upregulated in TM compared with HM.Chickens fed on TM diet had higher relative energy utilization compared to the HM birds.With increased corn storage,the relative digesta viscosity decreased significantly(P≤0.05).The total number of goblet cells and lymphocytes was lower in chickens fed the TM diet.The microbiota data showed that the TM chickens had decreased abundance of diarrheal bacteria such as Hungatella hathewayi and Bacteroides fragilis,and increased butyrate-producing bacteria such as Alistipes compared to the HM chickens.Conclusions:Storage of newly harvested corn induced the synthetic reaction of large molecules and changed the solubility of starch and protein with increasing soluble sugars and decreasing pasting properties that may improve the fermentation of intestinal microbiota,improve the energy utilization and protect gut health without the risk of diarrhea.