The search for a natural antimicrobial agent is ongoing and critical because of the rise and rapid proliferation of antibiotic-resistant pathogenic bacteria. The current study aims to examine the effect of Paenibacillus polymyxa AM20 as an alternative antibiotic and feed additive on Indian river broiler performance, digestive enzymes, thyroid hormones, lipid profile, hepatosomatic index, immunological response, gut bacteria, and antioxidant parameters. The bacterial isolate AM20 was identified at the gene level by isolating DNA and using PCR to detect genes. Based on 16S rRNA gene sequence analysis, the bacterial isolate was identified as Paenibacillus polymyxa. One hundred twenty Indian river broilers (1-day old) were randomly divided into 4 groups of 10 chicks each, with 3 replicates. The control group was fed a basal diet only, while the other 3 were administered control diets supplemented with P. polymyxa at 3 concentrations: 0.5, 1, and 1.5 mg/kg. The findings revealed that all groups that received graded amounts of P. polymyxa increased all growth parameters throughout the study. P. polymyxa treatment at 1.5 mg/kg increased body gain by 9% compared to the control due to increased feed intake (P = 0.0001), growth rate (P = 0.0001), and decreased feed conversion ratio. Compared to the control group, P. polymyxa (1.5 mg/kg) enhanced kidney functions in chickens by reducing uric acid and creatinine levels (P = 0.0451). Compared to the control group, alanine aminotransferase and aspartate transaminase levels in the liver were significantly reduced at all P. polymyxa doses. Liver function values were highest for P. polymyxa at 1.5 mg/kg. Compared to the control group, those whose diets included P. polymyxa had significantly better blood cholesterol levels, high-density lipoprotein, low-density lipoprotein, immunological response, thyroid function, and gut microbiota. In general, broiler chickens' economic efficiency was improved by including P. polymyxa in their diet, which also improved their growth performance, carcass dressing, specific blood biochemical levels and enzymes, and the composition of the gut microbiota.

Optimal dietary non-phytate phosphorus (NPP) is essential in poultry to maximise productive and reproductive
performance, along with indices of egg and bone quality. This study aimed to establish the NPP
requirements of egg-type duck breeders aged from 54 to 80 weeks on the following traits: egg production,
egg incubation, egg quality, tibial characteristics, reproductive organ, plasma indices, and the
expression of genes related to phosphorus absorption. Longyan duck breeders aged 54 weeks (n = 300)
were randomly allotted to five treatments, each containing six replicates of 10 individually caged birds.
Birds were fed corn-soybean meal-based diets containing 0.18, 0.25, 0.32, 0.38, and 0.45% NPP/kg for
27 weeks. The tested dietary NPP levels did not affect egg production or egg quality indices. The hatchling
weight of ducklings increased (quadratic, P < 0.01) as dietary NPP level increased, and the highest value
occurred with 0.25% NPP. The number of large yellow follicles (LYF), and the relative weights of LYF and
ovary showed linear and quadratic responses to dietary NPP levels; the lowest number and relative
weight of LYF occurred with 0.38% NPP, and the lowest ovarian weight was obtained with 0.25% NPP.
There were no differences in tibial length, breaking strength, and mineral density in response to dietary
NPP levels. In contrast, tibial content of Ca increased (linear, P < 0.01) with dietary NPP levels increasing
from 0.18 to 0.45%, and the tibial content of P increased at 0.32% NPP and the higher dietary NPP levels.
Plasma concentration of P showed a quadratic (P < 0.05) response to the dietary NPP levels, where the
highest value was seen at 0.38% NPP. In conclusion, dietary NPP levels from 0.18 to 0.45% had no effects
on egg production, and egg and tibial quality of duck breeders. The duck breeders fed a diet with 0.25%
NPP showed the highest hatchling weight of their offspring, while those fed 0.38% NPP had the lowest
number and relative weight of LYF. These results indicated that the diet with 0.25% NPP can be used
in egg-type duck breeders to improve the hatchling weight of their offspring, without adverse effects
on their productivity. The regression model indicated that the maximal hatchling weight of ducklings
was obtained from duck breeders fed the diet with 0.30% NPP.

Thepresentstudyinvestigatedtheinflu-enceofexposingquaileggstolow-dosegammaradiation(GR)andinovofeedingwith2sourcesofamixtureoftraceelements(Zn,Fe,andCu),includingsulfate(TES)andloadedwithmontmorillonite(TEM),onembryonicdevelopmentactivitiesandprehatchquality.Atotalof960eggsontheseventhdayofincubationwererandomlydividedinto6groups(160eggs/group)with4replicateof40eggsineach.A3£2factorialarrangementexperimentwasperformedandincluded3sourcesinovofeedingwithamixtureoftraceelements(Zn,Fe,andCu),including0mg/egg,50mgTES/egg,and50mgTEM/eggwitheggirradiationusing0and0.2GyfromGR.Eggsinjectedwith50mgTEM/eggandexposedto0.2GyfromGR(TEM/GR)wassignificantly(P≤0.05and0.01)higherinhatchability,hatchbodyweight,andrelativeorganweight(liver,gizzard,proventriculus,heart,andintes-tine).Theobtainedresultsindicatedsignificant(P≤0.05)decreasedintheserumconcentrationofmalondialdehyde(MDA)inTEM/GRgroup.Therewassignificant(P≤0.05)increasedofcatalase(CAT)activityandthe concentrationsofgrowthhormone(GH)andinsulin-likegrowthfactor-1(IGF-1)inTEM/GRgroup;however;totalantioxidantcapacity(T-AOC)wassignificant(P≤0.05)increasedinCT/GRgroup.SerumconcentrationsofimmunoglobulinM(IgM)(P≤0.05)andtumornecrosisfactor-alpha(TNF-a)wereincreasedintheTEM/CRgroup;theconcentrationoftransforminggrowthfactorbeta(TGF-b)significant(P≤0.05)increasedintheTEM/GRgroup;andinterleukins(IL6andIL10)showednosignificantdifferencesamongthegroups.Ourresultsshowedincreaseinthyroxineandmyostatincon-centrationswithTES/CRandCT/GRofourstudygroups,respectively.TherelativemRNAexpressionlevelsoftheGH,IGF-1,andFascellsurfacedeathreceptor(FAS)genesweresignificantly(P≤0.05and0.01)upre-gulatedinthelivertissueoftheTEM/GRgroupcom-paredwiththeothergroups.Inconclusion,TEM/GRwasthebesttreatmentforimprovingprehatchquality,increasingserumantioxidantenzymeactivities,andpro-motingtheexpressionofgrowthandimmunegenesinfer-tilizedquaileggs.

Intestinal oxidative stress in broilers is produced by chronic heat stress (HS) and has a negative
impact on poultry performance as it induces intestinal inflammation and promotes the invasion of
gram-negative bacteria, such as bacterial lipopolysaccharide (LPS). Therefore, dietary inclusion
of the antioxidant compound, ethoxyquin (EQ), could improve enteric antioxidant capacity,
immune responses, and the epithelial barrier, and maintain the symbiotic gut microbiota
community. To investigate the effects of EQ supplementation on alleviating enteric oxidative
stress in heat-stressed broilers, 200 one-day-old male Ross 308 broilers were randomly assigned
to four groups (n = 50 chicks/group; n = 10 chicks/replicate) and fed a basal diet supplemented
with 0 (CT), 50 (EQ-50), 100 (EQ-100), and 200 (EQ-200) mg EQ/ kg-1 for 5 weeks. The chicks
were raised in floor pens inside the broiler farm at a temperature and humidity index (THI) of 29
from day 21 to day 35. Growth performance traits, relative organ index, hepatic antioxidant
enzymes, serum immunity, total adenylate, and cytokine activities were improved in the EQ-50
group (linear or quadratic P < 0.05), promoting the relative mRNA expression of cytokine generelated
anti-inflammatory and growth factors. A distinct microbial community colonised the gut
microbiota in the EQ-50 group, with a high relative abundance of Lactobacillus,
Ligilactobacillus, Limosilactobacillus, Pediococcus, Blautia, and Faecalibacterium compared to
the other groups. Dietary supplementation with 50 mg EQ/ kg-1 for 5 wk attenuates enteric
oxidative stress and intestinal inflammation by enhancing serum immune and cytokine content
(IgG, IL-6, and TGF-β,) and symbiotic microbiota in heat-stressed broilers. EQ promotes the
expression of Hsp70, SOD2, GPx 4, IL-6, and IGF-1 cytokine gene-related anti-inflammatory and
growth factors in heat-stressed hepatic broilers. Collectively, EQ-50 could be a suitable feed
supplement for attenuating enteric oxidative stress and intestinal inflammation, thereby promoting
the productivity of heat-stressed broilers.

Abstract: Fusarium graminearum produces zearalenone (ZEA), a mycotoxin that is widely found in
food and feed products and is toxic to humans and livestock. Piper sarmentosum extract (PSE) inhibits
F. graminearum, and Oroxylin A appears to be a major antifungal compound in PSE. The aim of this
study is to quantify the Oroxylin A content in PSE using UPLC-QTOF-MS/MS, and to investigate the
antagonistic activity of Oroxylin A against F. graminearum and its inhibitory effect on ZEA production.
The results indicate that Oroxylin A inhibits both fungal growth and ZEA production in a dosedependent
manner. Oroxylin A treatment downregulated the mRNA expression of zearalenone
biosynthesis protein 1 (ZEB1) and zearalenone biosynthesis protein 2 (ZEB2). The metabolomics
analysis of F. graminearum mycelia indicated that the level of ribose 5-phosphate (R5P) deceased
(p < 0.05) after Oroxylin A treatment (64–128 ng/mL). Moreover, as the Oroxylin A treatment content
increased from 64 to 128 ng/mL, the levels of cis-aconitate (p < 0.05) and fumarate (p < 0.01) were
upregulated successively. A correlation analysis further showed that the decreased R5P level was
positively correlated with ZEB1 and ZEB2 expression, while the increased cis-aconitate and fumarate
levels were negatively correlated with ZEB1 and ZEB2 expression. These findings demonstrate the
potential of Oroxylin A as a natural agent to control toxigenic fungi and their mycotoxi

There is scarce information on the effects of feed storage period on poultry performance, which has not been investigated thus far. The aim of this study was to evaluate the effect of feeding broilers on feeds stored for different periods, with or without supplemental amino acids and vitamins. This study followed a factorial design (3 x 3), i.e. three dietary treatments and three feed storage periods. The three dietary treatments consisted of 1) a basal diet, 2) a basal diet including additional 5g methionine and 13 g lysine/kg, and 3) a basal diet including 3 g vitamin premix/kg. The 3 dietary treatments were tested with feeds stored for three different periods (Fresh, 4-months, and 6-months). A total of 450 one-day old broiler chicks were randomly assigned to the 9 treatments, each treatment contained 5 replicates of 10 chicks each (n= 50/group). The experiment lasted for 42 days. The results indicated that the final BWs of the birds received 4- or 6-months stored feed were greater than that of those fed fresh diets. The best total BWG was seen in the 4-month-stored feed group. There was no significant effect of storage period on total FC and FCR during period from 0 to 6 weeks of age, but the FCR tended to be better with the feed stored for 4 months. Haemoglobin was significantly higher in the birds fed fresh or 4-month stored feed. The percent of liver and spleen of the birds received 6-month-stored feed were the highest (P ˂ 0.01). Plasma content of albumin was higher in the birds received 4-or 6-month stored feed than the fresh diet. The use of additional amino acids increased the final BW and total BWG than those of the control and vitamin-supplemented treatments. The main effect of dietary treatments on percentages of immune organ weight was insignificant. The control (basal diet) recorded higher lymphocytes percentage, while T2 (BD + double level of amino acids) recorded higher eosinophil, heterophil and H/L ratio, whereas T3 (BD + double level of vitamins) recorded higher levels of haemoglobin, eosinophil, and monocytes. The main effect of dietary treatments on blood plasma proteins was insignificant. The tested treatments did not show a significant effect on broilers mortality. The detected values of AF1, AF2, AG1, and AG2 were very low in all 9 diets. In conclusion, enrichment of broiler feeds with higher levels of lysine and methionine (double level of NRC, 1994) together with antioxidants supplement could prolong their shelf-life to 6 months without any deleterious impact on productive performance of broiler chickens.

Abstract: In the poultry industry, there is an urgent need to evaluate and introduce natural, effective,
and safe alternatives for synthetic antibiotics, which have been banned in most countries. The present
study aimed to investigate the effects of dietary supplementation with Piper sarmentosum extract (PSE)
on the growth performance, intestinal barrier function, and metabolism of growing chickens. A total
of 400 seven-day-old female chicks were randomly assigned to four dietary treatments, each of which
consisted of five replicates and twenty birds each. The four experimental treatments were fed a basal
diet containing 0, 100, 200, and 300 mg PSE/kg (BC, PSE1, PSE2, and PSE3 groups), respectively.
The experiment lasted for 28 days. The results showed that dietary supplementation with PSE had
no significant effects on the final body weight, average daily gain (ADG), average daily feed intake
(ADFI), and the ratio of ADFI to ADG (F/G) (p > 0.05). Compared with the BC group, dietary
supplementation with 200–300 mg/kg PSE increased the villus height in the jejunum and ileum of
chickens (p < 0.05). The PSE-treated groups significantly increased the mRNA expression of Occludin,
ZO-1, and Claudin-1 in the ileal mucosa of chickens (p < 0.05). In addition, a significant decrease in
ileal TNF- and IL-8 mRNA expression (p < 0.05) and a significant increase in IL-22 (p < 0.05) were
observed in the PSE2 treatment compared to the BC group. Additionally, three gut metabolites (i.e.,
citrate, isocitrate, and spermine) showed significant differences among treatments (p < 0.05) and
were involved in the tricarboxylic acid (TCA) cycle, the transfer of acetyl groups into mitochondria,
and spermidine and spermine biosynthesis, respectively. In conclusion, the findings obtained here
indicate that supplemental PSE can enhance the anti-inflammatory capacity and intestinal mucosal
barrier function of chickens.

The access of vinasse leachates to water bodies and groundwater exacerbates environmental problems, especially eutrophication. Therefore, a column experiment was performed to examine the effect of adding zeolite (ZL), bone char (BC), and wood chips biochar (WCB) in the presence of vinasse on carbon dioxide (CO2) emission, leaching dissolved organic carbon (DOC) and ammonium (NH4+) in calcareous sandy soil, as well as studying the kinetics of leaching dissolved organic carbon and ammonium. This column experiment contains four treatments: soil alone (CK), soil + zeolite (SZL), soil + bone char (SBC), and soil + wood chips biochar (SWCB). These amendments were applied to the soil at a level of 4%. Vinasse was added to all treatments at a level of 13 mL per column. The leached total cumulative DOC and total cumulative soluble ammonium amounts decreased significantly with applying ZL, BC, and WCB compared with the soil alone. The effectiveness of these amendments in lowering the total cumulative DOC leaching is in the order of SBC > SWCB > SZL >CK. However, the effectiveness of these amendments in decreasing the total cumulative NH4+ leaching is in the order of SZL > SWCB > SBC >CK. The rate constant (k) of DOC leaching decreased significantly with the application of bone char compared to soil alone treatment. In the presence of vinasse, the apparent half‑life of leached DOC from the soil was 8.1, 12.9, 36.7, and 15.5 days for soil CK, SZL, SBC, and SWCB treatments, respectively. Half‑life values of leached soluble ammonium from the soil in the presence of vinasse addition were 10.1, 39.5, 28.5, and 37.9 days for CK, SZL, SBC, and SWCB treatments, respectively. Amending soil with BC increased significantly the phosphorus availability, however, applying ZL and BC caused a significant increase in the available potassium in calcareous sandy soil compared to the control treatment. According to these results, it is recommended not to add vinasse alone to sandy soils, but it is preferred to be co‑applied with BC amendment at the level of 4% better than ZL and WCB. This would decrease leaching DOC and ammonium to the water table and groundwater as well as enhance nutrient retention in the soil, which in turn, plays a vital role in reducing the harmful effect of vinasse and improving soil fertility.
 

Pollution by petroleum hydrocarbons (PHCs) is a current environmental challenge all over the world. Therefore, this study was established to evaluate the impact of calotropis procera biochar (CPB) produced at various pyrolysis temperatures on carbon emission, ammonia volatilization, degradation of PHCs, and properties of artificial petroleum-contaminated soil. An incubation experiment was performed with CPB produced at 400 °C (CPB400) and 650 °C (CPB650). The artificial petroleum-contaminated soil was amended with biochar at doses of 1, 2.5, and 5% along with the unamended soil (control) and incubated for 78 days. The results demonstrate that the CO₂-C emission rate in petroleum-contaminated soil increases significantly with all biochar applications compared to the control treatment. Adding CPB400 into the soil, at all doses, decreased cumulative NH₃ emission significantly, in comparison with control and CPB650 treatments during all incubation periods. Total PHCs concentration in the soil reduced significantly with CPB400 application at 2.5% compared to the control treatment. CPB400 was more effective in declining total PHCs content in the soil than biochar produced at 650 °C. Available phosphorus and potassium in petroleum-contaminated soil increased significantly under biochar applications compared with the control treatment. Using Calotropis procera biochar produced by low-temperature pyrolysis as an amendment can play a vital role in the remediation of PHCs in polluted soils, it is also a promising way to improve the quality indicators of petroleum-contaminated soil.