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Abstract Eimeria spp. are the pathogen that causes coccidiosis, a significant disease that affects intensively reared livestock, especially poultry. Anticoccidial feed additives, chemicals, and ionophores have routinely been employed to reduce Eimeria infections in broiler production. Therefore, the shift to antibiotic-free and organic farming necessitates novel coccidiosis preventive strategies. The present study evaluated the effects of potential feed additives, liver free and chitosan, against Eimeria tenella infection in White Leghorn broiler female chickens. One hundred sixty-five 1-day-old White Leghorn broiler female chicks were divided into 11 groups (15 female chicks per group), including the positive control group (G1), the negative control group (G2), a chitosan-treated group (G3), a chitosan-treated-infected group (G4), the liver free-treated group (G5), the liver free-treated-infected group (G6), the liver free-and-chitosan-treated group (G7), the liver free-and-chitosan-infected group (G8), the therapeutic liver free-and-chitosan-treated-infected group (G9), the sulfaquinoxaline-treated group (G10), and the sulfaquinoxaline-treated-infected group (G11). Chitosan was fed to the chicks in G3 and G4 as a preventative measure at a dose of 250 mg/kg. The G5 and G6 groups received 1.5 mg/kg of Liverfree. The G7 and G8 groups received chitosan and Liverfree. The G10 and G11 groups were administered 2 g/L of sulfaquinoxaline. From the moment the chicks arrived at Foshan University (one-day-old chicks) until the completion of the experiment, all medications were given to them as a preventative measure. G8 did; however, receive chitosan and liver free as therapeutic supplements at 7 dpi. The current study showed that the combination of liver free and chitosan can achieve better prophylactic and therapeutic effects than either alone. In E. tenella challenged chickens, G8 and G9 chickens showed reduced oocyst shedding and lesion score, improved growth performance (body weight, body weight gain, feed intake, feed conversion ratio, and mortality rate), and cecal histology. The current study demonstrates that combining liver free and chitosan has superior preventive and therapeutic benefits than either alone, and they could also be used as alternative anticoccidial agents.

ABSTRACT: Intensive broiler production systems face challenges like enteric diseases, impacting global food security. Strategies to enhance broiler immunity and gut health, particularly amidst antibiotic growth promoter restrictions, are crucial. The present study investigated the combined effects of fenugreek seeds (FS) and Bacillus-based direct-fed microbials (DFM) on immune-related gene expression in the ileum and alteration of microbial population in the cecum of broiler. The study involved 160 Ross 308 broiler chicks, which were divided into four groups consisting of 5 replicates, each containing eight birds. The chicks were grown for a period of 42 d, during which they had ad libitum access to feed and water. Dietary treatments were: Control (basal diet), FS5 (basal + 5g/kg fenugreek seeds), FS5DFM (basal + 5g/kg fenugreek seeds + 0.1g/kg Bacillus-based DFM), and DFM (basal + 0.1g/kg Bacillus-based DFM). Ileum tissue and cecal contents were collected on d 42 for gene expression and gut microbiome analysis. Ileal gene expression analysis revealed the downregulation of IL-6, IL-8L2, CASP6, PTGS2, and IRF7 in both FSs and DFMs groups compared to the control, suggesting individual immunomodulatory effects. However, avian β-defensin genes exhibited complex regulation, highlighting the need for further investigation. Cecal microbiome diversity remained stable, with subtle shifts in specific taxa influenced by FSs and DFMs. Interestingly, the combination of the FSs and DFMs uniquely impacted specific taxa, including Clostridiales vadin BB60. These findings suggest that both FSs and DFMs demonstrated potential for improving broiler immunity through inflammation reduction. The combination of FSs and DFMs offers a synergistic effect in immune modulation and specific microbial modulation, warranting further investigation with pathogen challenge models for comprehensive understanding.

ABSTRACT: This study explored the ability of formic acid (FA) to replace antibiotics in broiler chicken diets. It examined how FA affected the chickens' growth, carcass characteristics, blood chemistry, and gut bacteria. The experiment randomly assigned 300 one-day-old (Ross 308) broiler chicks to 5 groups, each divided into 6 replicates with 10 unsexed chicks. The following were the treatments: 1st group, negative control (NC): only received a basal diet; 2nd group, positive control (PC): received a basal diet supplemented with 0.5 grams of Colistin antibiotic per kilogram of diet; 3rd, 4th, and 5th groups (FA2, FA4, and FA6) these groups received a basal diet along with formic acid added at increasing levels: 2, 4, and 6 Cm3 per kilogram of diet, respectively. Results found no significant differences in live body weight (LBW) or body weight gain (BWG) between treatment groups, except for LBW at one week and BWG at 0 to 1 and 4 to 5 wk of age. No significant variations were found in feed intake (FI) and feed conversion ratio (FCR) among the treatment groups, excluding FI and FCR at 1 to 2 wk of age. The treatments significantly impacted carcass traits, dressing percentage, breast meat, thigh meat, spleen, giblets, blood levels of urea, creatinine, total protein, globulin, and albumin, as well as the activity of enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in chicks fed different diets compared to control groups. The addition of FA to the diet significantly impacted antioxidant levels. Also, the FA2 group had the highest total bacterial count (TBC). However, the FA6 group was the opposite; it had the lowest levels of harmful bacteria, such as E. coli and Coliform. Supplementing broiler diets with formic acid improves blood parameters, antioxidant activity, and gut bacteria counts, with 4.0 cm³ formic acid/kg diet supplementation promoting optimal broiler health and product quality.

ABSTRACT: Cellulitis is an important disease in commercial turkey farms associated with significant economic loss. Although the etiology of cellulitis is not fully elucidated, Clostridium septicum (C. septicum) is one of the main causes of this infectious disease. In this study, we report the development of a quantitative real-time PCR (qRT PCR) assay targeting the alpha-toxin gene (csa), which involves a prior 15-cyle PCR using a nested pair of primers to increase the detection sensitivity. Additionally, the TaqMan probe was employed to increase the target-specificity of the assay. The performance of our nested qRT-PCR assay was evaluated using Clostridium isolates from turkey farms, representing both septicum and non-septicum species, as well as sponge swab samples from turkey farms. Our step-by-step development of the assay showed that the csa gene is a suitable target for specific detection of C. septicum strains and that the inclusion of nested PCR step significantly increased the detection sensitivity of the final qRT PCR assay. The performance of the assay was also validated by a high correlation of the threshold cycle numbers of the qRT PCR assay with the relative abundance of C. septicum read counts in 16S rRNA gene microbiota profiles of the C. septicum-containing samples from turkey farms.

ABSTRACT: This experiment was carried out to investigate the nutritional value of Spirulina and Dunaliella (SD) combination levels (0, 0.5, 1.0, 1.5, and 2.0 g/kg) that affected the laying Japanese quail's efficiency, egg quality, fertility, and blood biological indicators. A total of 150 adult Japanese quails, aged 8 wk, were divided into 5 treatments at random, each consisting of 30 quails. There were 5 duplicates for every treatment, with 2 male and 4 female quails in each. Comparing the addition of various concentrations of a mixture of SD to the control treatment, the results showed no substantial rise in egg production, egg weight, or egg mass. When compared to the control group, final body weight (FBW) was improved with SD supplementation. The quails in control consumed more feed intake (FI) (p < 0.05), and they were different from the groups who got SD therapy in that they had a regular feed conversion ratio (FCR). The percentages of hatchability and fertility increased when SD was added to quail meals at up to 1.00 g/kg. When compared to the control quail, the quail supplemented with SD levels showed a non-significant rise in albumin%, yolk%, Haugh unit, and unit surface shell weight (USSW), as well as an increase in eggshell percentage and a drop in egg shape index (p < 0.05). Renal and hepatic enzyme functioning improved when SD was added to the diets. Additionally, lipid profile indicators were reduced by SD supplementation (except low-density lipoprotein-LDL). Moreover, compared to the control, incorporating SD led to a nonsignificant rise in immunoglobulin concentrations (IgG and IgM). In conclusion, adding SD to the diet can improve body weight, lipid profile, immunological response, and liver and kidney functions in Japanese quail.

ABSTRACT: The purpose of this investigation was to evaluate the impacts of vitamin A (VA) supplementation in feed at levels of 0 (control), 2,000, 4,000, 6,000, and 8,000 IU VA/kg diet on the reproductive efficiency and antioxidative properties of aged Sinai laying hens at 52 wk of age (n = 300 females and 30 males) in 6 replicates (10 females + 1 male/replicate). As well as blood biochemical indicators, carcass characteristics, growth performance, immunity, and the antioxidative status of their chicks. Results showed that diets supplemented with 2,000 or 6,000 IU/kg of VA increased fertility rate and decreased early embryonic mortality (P < 0.05). Increasing VA from 4,000 to 6,000 IU/kg significantly boosted hatchability rates. All VA levels significantly enhanced glutathione peroxidase (GPx) and reduced malondialdehyde (MDA) and late embryonic mortality. In the shell gland, dietary supplementation of 6,000 or 8,000 IU/kg of VA enhanced actions of GPx actions, catalase (CAT), and superoxide dismutase (SOD). In hatched chicks, all VA levels boosted (P < 0.05) hemoglobin, red blood cell count, and serum concentration of total proteins and IgA while decreasing eosinophils percentage and aspartate aminotransferase activity (AST) concentration. Dietary VA supplementations from 4,000 to 8,000 IU/kg improved lymphocytes, serum total antioxidant capacity (TAC), SOD, and IgM, while decreasing heterophils, heterophils/lymphocytes ratio, and creatinine in hatched chicks. Serum triglyceride concentration was reduced by adding 6,000 or 8,000 IU/kg of VA, while globulin and high-density lipoprotein concentrations were heightened only by 8,000 IU/kg of VA. It could be concluded that the dietary supplementation of VA (6,000 IU/kg) improved reproductive efficiency and antioxidative status in the liver and the shell gland of aged laying hens and improved hemato-biochemicals parameters, antioxidative status, and immunity of their offspring.

ABSTRACT: This study used 300 1-day-old, sexless, developing chicks of Japanese quail to estimate the ability of vitamin C and/or garlic to antagonize the venomous influence of cadmium (Cd) on the hematological, immunological, and performance characteristics of developing Japanese quail. The quail was separated into 5 similar groups of 60 chicks apiece, and 6 duplicates (10 each) were given to each sub-group. The control group received a basal diet without any supplements. The Cd group was nourished with a basal diet of + 80 mg cadmium chloride (CdCl2)/kg diet. The 3rd group was fed a basal diet + 80 mg CdCl2/kg diet and complemented with a 200 mg Vitamin C (Cd + C)/kg diet. The 4th group was nourished with a basal diet + 80 mg CdCl2/kg diet and complemented by a 500 mg dried garlic powder (Cd + G)/kg diet. The 5th group was fed a basal diet + 80 mg CdCl2/kg diet, complemented by a 200 mg vitamin C/kg diet + 500 mg dried garlic powder (Cd + CG)/kg diet. Results showed that in the 5th group in which cadmium was added together with Vit C + garlic, there was an improvement in both live weight gain (1–42 d) and feed consumption (1–21 and 1–42 d ) compared to the group in which Cd was added alone. The addition of Vit C alone and together with garlic seems to completely improve the cadmium-related increase in alkaline phosphatase (ALP), and Aspartate aminotransferase (AST), and Malondialdehyde (MDA) levels when compared to the control. Compared to cadmium-polluted diets, quail that got cadmium and feed additives significantly reduced cadmium residue. In addition, the cadmium group's serum immunoglobulin M (IgM) level decreased significantly. These data imply that dietary supplementation with (C) or (G) may be beneficial in retrogressing the drop in immunoglobulin G (IgG) and IgM caused by Cd and minimizing Cd's deleterious influence on immunity.

ABSTRACT: Escherichia coli (E. coli) is a significant challenge in the poultry industry due to their related use of antimicrobial compounds and the drastic losses in production and livability. This study investigated the preventive impacts of dietary supplementation of Salvadora persica (SP) and/or Pulicaria undulata (PU) extracts on growth traits, biochemical and immune parameters, and related gene expression of E. coli-infected broilers. A total of 120 one-day-old Cobb broilers were used. The chicks were allocated into eight equal groups (3 replicates/ group; 5 chicks per each replicate) as follows: G1; control negative, G2; SP-treated, G3; PU-treated, G4; SP/PU-treated, G5; E. coli infected, G6; E. coli infected and SP-treated, G7; E. coli infected and PU-treated, G8; E. coli infected and SP/PU-treated groups. Results revealed significant improvement in average body weight (ABW), average weight gain (AWG) and feed conversion ratio (FCR) in broilers fed diets supplemented with SP and/or PU compared to control and E. coli infected groups. Moreover, significant (P < 0.05) reduction in ALT, AST, creatinine, and uric acid was reported in other treated groups compared to the single E. coli-infected broilers. On the contrary, a significant increase (P < 0.05) in serum immunoglobulin and protein concentration was also reported in treated groups when compared to E. coli-infected untreated group. In addition, feeding broilers with SP and/or PU significantly improved (P < 0.05) the relative weight of immune-related organs and gene expression of TLR-15, with subsequent down-regulation of IL-1β and TNF-α mRNA transcripts. Supplementing broilers with dietary SP and/or PU could be promising in the prevention of E. coli infection via stimulating significant improvement of immune-related gene expression, immune-related organ weight, and down-regulation of inflammatory-related genes, with subsequent enhancement of the growth performance of broiler chickens.

ABSTRACT: The aim of this research was to evaluate the influence of lettuce seed oil (LSO) on the performance, carcass yield, kidney and liver indices, immunity, lipid profile, and cecal microbiota of fattening chicks. A total of 200, 7-day-old Cobb-500 were distributed into 5 experimental groups; each group contained 5 replicates with 8 birds each. The first group 1) the basal diet (only); 2) the basal diet plus lettuce seed oil (0.50 mL/kg); 3) the basal diet plus lettuce seed oil (1.00 mL/kg); 4) the basal diet plus lettuce seed oil (1.50 mL/kg); and 5) the basal diet plus lettuce seed oil (2.00 mL/kg). No significant effect was observed on growth performance, carcass traits, or kidney function at any level of oil. But, liver function was significantly affected due to LSO levels. Serum lipid profiles (total cholesterol—TC, triglyceride—TG, low-density lipoprotein—LDL, and very low-density lipoprotein—VLDL) were significantly reduced by using LSO levels compared to the control group. Dietary LSO significantly increased immunological and antioxidant parameters, except for malondialdehyde-MDA, which was reduced. On the other hand, the cecal microbiota was significantly improved by LSO additives. It was concluded that the dietary supplementation of LSO had beneficial effects on liver and kidney functions, lipid profile, immunity, antioxidant parameters, and the bacteriology of fattening chicks.

ABSTRACT: The investigation examined the use of willow leaf extract (WLE) on broiler chickens, examining carcass characteristics, cecal microbiota, antioxidants, and blood parameters. In 4 groups of 300 chicks, a basal diet was given for 5 wk, and the first treatment was basal diet (C). The diets for the remaining 3 treatments (WLE150, WLE300, and WLE450) contained 150, 300, and 450 mg of willow leaf extract /kg, respectively. The study found that birds fed willow leaf extract supplements had significantly greater body weight (BW), body weight gain (BWG), and enhanced feed conversion ratio (FCR) vs. the control group. Birds fed at 450 mg/kg food showed the greatest growth features, carcass weight, liver weight, lower abdominal fat, better low-density lipoprotein (LDL), and high-density lipoprotein (HDL) concentrations, and highest hematological characteristics. Chickens fed diets supplemented with varied doses of willow leaf extract showed significantly increased antioxidant enzyme activity, with higher amounts of glutathione peroxidase (GPx) activity, superoxide dismutase (SOD), total antioxidant capacity (TAC), and lower malondialdehyde (MDA). However, in the study, birds fed a diet supplemented with 450 mg of willow leaf extract per kg meal showed a significant drop of 13.02%, which found no significant variations in hazardous bacteria (Escherichia coli) across 2 treatments (WLE150 and WLE300). In addition, the study discovered that birds fed with varied doses of willow leaf extract had fewer cecum infections (Staphylococci aureus). We conclude that using willow at a level of 450 mg/kg diet can significantly enhance the BWG, FCR, antioxidant levels and beneficial bacteria activity besides the condition of broiler chicken's general health.

ABSTRACT: Olive oil (OL) production is the most significant agro-industrial business and has a high impact on the economy of numerous Mediterranean countries. However, OL extraction results in massive amounts of byproducts, including a solid residue (olive cake or olive pomace) and an aqueous stage (olive mill wastewater), which have serious environmental effects due to their hazardous nature and excessive organic content. Despite these byproducts causing environmental pollution, they can be applied for animal feeding. According to the literature, OL or its derivatives have been used to promote broiler performance, feed utilization, and health status in broilers as growth promoters or protein sources. Furthermore, using OL and its derivatives could improve heat resistance in stressed broilers via struggling thermal stress effects. In this framework, we highlighted the use of OL and its byproducts in broiler feeding to promote performance and health status. Additionally, the role of these byproducts and OL in combating thermal stress is investigated for sustainable strategy and promoting circular economy in broiler industry.

ABSTRACT: The present study aimed to investigate the impact of various concentrations of ginger and cinnamon oils as antibiotic substitutes on some blood biochemical parameters, antioxidant capacity, and histopathological profile of the liver and gut of growing Japanese. A total of 900 Japanese quails were randomly allotted into 6 treatment groups. Each group had 5 replicates (30 chicks each). The first group received a basal diet and served as the control, while the second received a basal diet plus 0.5 g of colistin antibiotic/kg diet. The third and fourth groups were supplemented with 0.5 mL and 1.0 mL of ginger oil (GO)/kg diet, respectively. While the fifth and sixth groups received basal diet with 0.5 and 1.0 mL of cinnamon oil (CO)/kg diet, respectively. Results showed that adding herbal oils significantly (P < 0.05) decreased the aspartate aminotransferase (AST) and urea levels compared to control and colistin groups. Various levels of GO and CO significantly (P < 0.05) reduced cholesterol levels compared to control birds. Compared to the control and antibiotic groups, Japanese quails supplemented with various levels of herbal oils (GO and CO) had more extraordinarily significant (P < 0.05) values for total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GPX), and glutathione reductase (GSR). Regarding histopathologic examination, the jejunum displayed a nearly empty lumen, a few fusions, and mild goblet cell metaplasia. On the other hand, the duodenum looked tall and had a few fusions of villi and remnants of removal in its lumina. It could be concluded that cinnamon and GO improved birds’ blood biochemical parameters, electorate oxidative stress, and enhanced intestinal and hepatic histology of the treated quails. Also, the levels of 0.5 mL CO and 0.5 mL GO may be an acceptable substitute for antibiotics (colistin) in the diets of growing Japanese quail.

ABSTRACT: The current study aimed to determine the effects of different levels of fermented Juncao grass (FG) on growth parameters, blood constituents, immunity, and antioxidative properties of broilers. A total of 240 (21-d-old) broiler chicks were randomly distributed to four dietary treatments of sixty birds, with six replicate pens and ten birds in each. Fermented grass was added to the basal diet at four levels with 0, 5, 10, and 15% FG. The results revealed that broilers fed 5% FG had significantly higher (P < 0.05) final body weight (FBW), average daily gain (WG) and average daily feed intake (ADFI). The best conversion ratio (FCR) was recorded for broilers supplemented with 5% FG compared to the group supplemented with 15% FG (P < 0.05). Increasing FG % decreased (P < 0.05) anti-inflammatory cytokines IL-4, IL-6, and IL-10. However, FG increased (P < 0.05) proinflammatory cytokines IFN-γ, IL-1, IL-2, IL-12, and TNF-α (P < 0.05). Moreover, IgA, IgG, and IgM levels increased (P < 0.05) with increasing FG %. In addition, increasing FG % in broiler rations significantly increased (P < 0.05) serum antioxidant levels of T-AOC, GSH-PX, SOD, CAT, NO and GSH, but decreased (P < 0.05) MDA levels compared to the control group. Conclusively, fermented Juncao grass would be considered a novel herbal feed additive for improving broiler performance, immunity, antioxidant, and health status. Nevertheless, further research at the molecular level is needed to quantify the effects of these herbal components on cellular and humoral immune functions in broiler chickens.

ABSTRACT: For the evaluation of the effects of various quantities of crude protein (CP) and metabolizable energy (ME) (kcal/kg diet) in Japanese quail lay chicken meals on performance, digestion coefficients, and nutritional value. An experiment was undertaken in the summer season; 216 eight-wk-old Japanese quail laying hens were allocated at random to 1 of 9 groups in a factorial design (3 × 3) experiment involving 3 levels of CP (18, 20, and 22%) plus 3 levels of ME (2,800, 2,900, or 3,000 kcal/kg ME diet). The testing period lasted from 8 to 20 wk. The findings demonstrated that the relationship between protein and energy levels significantly impacted all productive outcomes except the feed conversion ratio (P < 0.01). Throughout the overall period 8 to 20 wk of age, the greatest measurements of egg weight (11.9 g) and egg mass (8.33 g) occurred in hens fed 20% CP with 2,900 kcal/kg diet, yet the lowest values of egg weight (11.2 g) and egg mass (6.72 g) were noticed in hens fed 22% CP with 2,900 kcal/kg diet. The data showed that the combination of dietary energy and protein levels had a substantial (P < 0.05 and P < 0.01) effect on all egg quality trials evaluated. The results indicated that the interaction between dietary energy and protein concentrations substantially impacted DM and EE digestion ratios (P < 0.01). Finally, when feeding layer Japanese quail between the ages of 8 and 20 wk during the summer, a dietary energy content of 2,900 kcal ME/kg with 20% CP is recommended.

The exploration for effective in-feed additives is growing owing to the global climatic change trend to alleviate the negative effects of heat stress in laying hens. This research assessed the potential of using B-glucan (G) as an antiheat stress agent in Matrouh laying hens subjected to early heat shock programs during the growing period. Factorial design (3 × 3) was used, including 3 levels of heat stress (control, heat shock at 3 d and at 3 d and 8 wk of age) and 3 levels of β-glucan (0, 100, and 200 mg β-glucan /kg diet). During the first 12 wk of egg production (EP), treatments were exposed to heat challenge. The results revealed that heat shock program applications at 3 d and 8 wk of age significantly decreased body weight at 36 wk of age (P < 0.05) and reduced (P < 0.05) feed intake (FI). While significantly (P < 0.05) improved feed conversion ratio (FCR), hemoglobin, RBCs, WBCs, immunoglobulin M (IgM), immunoglobulin G (IgG), and Heat shock protein (HSP70) of the Liver (P < 0.01) as compared with the control group. At the same time, there was a decrease in lymphocyte%, H/L ratio, cortisol, and T3 compared to the thermo-neutral control. When compared to the control group, hens fed a diet containing 200 mg of βG significantly (P < 0.05) improved body weight at 16 wk and final weight at 36 wk, feed conversion (FCR) (g. feed/g. egg mass), hen-day egg production, and egg mass, as well as the digestibility coefficients of crude protein (CP), dry matter (DM), metabolizable energy (ME), and cortisol. The interactions between heat chock programs and βG levels were nonsignificant for the most studied traits except daily feed intake. Therefore, the early heat shock exposure 2 times and supplementation of Β-glucan (βG) at 200 mg/kg diet during the growth period for laying hens that are exposed to heat stress during the reproductive period could improve productive, reproductive performance, HSP70 level and enhance immunity responses.

Heat stress (HS) is still the essential environmental agent influencing the poultry industry. Research on HS in poultry has progressively acquired growing interest because of increased attention to climate alteration. Poultry can survive at certain zone of environmental temperatures, so it could be considered homoeothermic. In poultry, the normal body temperature is essential to enhance the internal environment for growth, which is achieved by normal environmental temperature. Recently, many studies have revealed that HS could cause mitochondrial dysfunction in broilers by inducing redox dysfunction, increasing uncoupling protein, boosting lipid and protein oxidation, and oxidative stress. Moreover, HS diminished the energy suppliers supported by mitochondria activity. A novel strategy for combating the negative influences of HS via boosting the mitochondria function through enrichment of the diets with mitochondria enhancers was also described in this review. Finally, the current review highlights the mitochondria dysfunction induced by HS in broilers and attempts to boost mitochondria functionality by enriching mitochondria enhancers to broiler diets.

ABSTRACT: The present study aimed to investigate the efficacy of probiotics and prebiotics in controlling Escherichia coli (E. coli) spp. isolated from chicken. A total of 230 birds representing 19 different commercial breeds were taken from various points. Birds were monitored for postmortem and clinical investigation. Aseptically collected liver samples, lungs, kidneys, hearts, and yolk sacs were then subjected to bacterial isolation and identification. E. coli were observed in 9 pooled samples from 120 examined with an incidence of 7.5%. Nine farms were E. coli-positive, with an incidence of farm infection of 47.3%. The 9 suspected isolates of E. coli were profiled by morphological and microbiological identification of the colony, motility, and gram reaction. The serogroup analysis showed 9 different E. coli for which 3 other groups were identified: 2 E. coli O78, 3 E. coli O111, and 4 untyped groups. Nine isolates of E. coli were subjected to PCR. Molecular detection of 9 strains was conducted to find the virulence genes of E. coli strains (8 STX1, 4 STX2, and 9 EAE). Probiotics and prebiotics significantly increased the total erythrocytic and leukocytic counts throughout the experiment. The phagocytic percentage's main values at 14 d were 47 and 30%, respectively. An increase in the humoral immunity against Newcastle disease (ND) was noticed after ND vaccination. The geometric mean (HI) was 5.9 and 4.2 for probiotic and prebiotic, respectively. It could be concluded that probiotics and prebiotics could stimulate a nonspecific immune response against experimental infection with a virulent strain of E. coli spp.

ABSTRACT: This study investigated the impact of Arthrospira platensis (A. platensis) as a dietary supplement on growth, carcass criteria, liver and kidney function, lipid profile, and immunity of growing Japanese quails. In a 28-day experiment, 240 unsexed 7-day-old quail chicks were used. The quail chicks were divided into 4 treatment sets, each comprising 6 replicates and 10 quail chicks. Group (1) fed the basal diet with no supplements, group (2) fed the basal diet supplemented with 0.25 g A. platensis/kg diet, group (3) fed the basal diet supplemented with 0.50 g A. platensis/kg diet, and group (4) fed the basal diet supplemented with 1.00 g A. platensis/kg diet. The results of this study revealed that the birds that were fed 0.50 g. or 1.00 g A. platensis/kg diet had superior final body weights, body weight gains, feed conversion ratios, and carcass criteria compared to control, furthermore, had significant (P < 0.05) lower levels of liver enzymes and kidney function markers compared to control. Furthermore, birds fed 0.50 g or 1.00 g A. platensis/kg diet had significantly (P < 0.05) reduced cholesterol, triglycerides, and LDL cholesterol levels compared to control. Dietary supplementation of A. platensis at 0.50 and 1.00 g/kg diet significantly (P < 0.05) increased total antioxidant capacity, total immunoglobulin, and lysozyme activity levels. Using A. platensis at 0.5 or 1 g/kg diet levels during the Japanese quails' growth period can improve growth, carcass criteria, liver and kidney function, lipid profile, and immunity.

ABSTRACT: One of the most intriguing areas of research and innovation in the animal production and food sector recently has been designed-enriched products. These items are regarded as functional foods because they feature components that have advantageous physiological impacts on human health. In the production of poultry, designed eggs constitute a significant category of functional foods. The present study hypothesized that adding different kinds of oils to quail diets will help produce designer eggs rich in omega-3 and 6 fatty acids in addition to enhancing productive performance. So, this study examined how linseed (flaxseed) and canola oils with various levels can affect lipid metabolism, immune function, and the amount of n-3 polyunsaturated fatty acids (n-3 PUFA) in Japanese quail eggs. This work was conducted using 3 different vegetable oils (sunflower, linseed, and canola oils) and 3 different antioxidant supplements (0, 250 mg vitamin E/kg feed, and 1,000 mg ginger/kg feed) in a 3 × 3 factorial experiment. When linseed or canola oil was added to the diet, the number of fatty acids in the egg yolks of Japanese quail layers fell by (12.7 and 18.9%) and (41.4 and 24.6%), respectively. The amounts of saturated and monounsaturated fatty acids in total eggs fell by 21.9 and 14.6% and 24.5 and 15.8%, respectively, at 20 wk of age. However, when linseed and canola oil were added to the diet, the sum n-3 PUFA content in the egg yolk of Japanese quail-laying hens was noticeably raised at 15 and 20 wk of age. At 15 and 20 wk of age, the same groups' total n-6 PUFA content considerably increased compared to the group that did not receive flaxseed. In conclusion, during the laying period of Japanese quail, linseed oil, canola oil, vitamin E, or ginger positively affected productivity, blood hematology, constituents, resistance, lipid digestion system, and antioxidative properties in serum and egg yolk.

BackgroundThe objective of the present study was to evaluate the potential synergistic impact of the combination of fenugreek seeds (FS) and Bacillus-based direct-fed microbials (DFM) on growth performance, intestinal health, and hematological parameters of broiler chickens.MethodsA total of 160 one-day-old (Ross 308) broiler chicks were randomly assigned to a 2 × 2 factorial arrangement, with two levels of FS (0 and 5 g/kg) and two levels of Bacillus-DFM (0 and 0.1 g/kg), with five replicates of 8 birds each.ResultsThe result showed that dietary supplementation of FS at 5 g/kg did not improve the growth performance of broilers but impaired the early growth performance by reducing body weight gain and increasing feed conversion ratio, which was recovered during finisher phase. Dietary supplementation of Bacillus-based DFM at 0.1 g/kg did not affect the performance variables but increased the feed conversion ratio. The interaction of fenugreek seeds and Bacillus-based DFM showed synergistic effects on growth performance during the later stages of production. However, antagonistic effects were observed on the blood parameters and the gut morphology.ConclusionThis study demonstrated that FS and DFM had different effects on the broiler health and production depending on the phase of production. The interaction between FS and DFM revealed synergistic effects on growth performance during the finisher phase, but antagonistic effects on blood parameters and gut morphology. Further studies are needed to elucidate the underlying mechanisms and optimize the dosage and combination of FS and DFM for broiler health and production.

ABSTRACT: Enterococcus cecorum (EC) has been associated with septicemia and early mortality in broiler chickens. There is limited research investigating the pathogenicity of EC field strains obtained from affected birds. The purpose of this study was to evaluate the effect of in-ovo administration into the amnion with different EC field isolates at d 18 of embryogenesis (DOE18). In Exp 1, 7 EC field isolates alone or in combination (EC1–EC3, EC4–EC5, EC6, and EC7) were selected based on phenotypic characteristics and evaluated at different concentrations (1 × 102, 1 × 104, and 1 × 106 CFU/200 µL/embryo) to assess the impact on early performance and macroscopic lesions. Three isolates (n = 3; EC2, EC5, EC7) were selected for additional evaluation based on the significant (P < 0.05) BWG reduction (d 0–21) compared to the negative control (NC) and the presence of macroscopic lesions observed during posting sessions at d 14 and d 21. An additional isolate associated with enterococcal spondylitis was included in Exp 2 (EC11B). Treatment groups for Exp 2 include: 1) NC, 2) EC2, 3) EC5, 4) EC7, and 5) EC11B (n = 90–120/embryos/group). Groups 2 to 5 were challenged at 1 × 102 CFU/200 µL/embryo by in-ovo injection into the amnion at DOE18. Chicks were placed in battery cages for the duration of the study (21 d), and pen weights were recorded at d 0, d 7, d 14, and d 21 to calculate average BW and BWG. At d 14 and d 21 posthatch, liver, spleen, free thoracic vertebrae (FTV), and femoral head (FH) were aseptically collected to enumerate Enterococcus spp. using Chromagar Orientation as the selective media. Cecal contents were collected at d 21 to evaluate the effect of EC challenge on the cecal microbiome composition. There was a significant (P < 0.05) reduction in BW at d 21, and BWG from d 14 to 21 and d 0 to 21, for EC7 and EC11B. Enterococcus cecorum was recovered from the FTV of all challenged groups at d 14 and d 21. The most representative lesions were pericarditis, hydropericardium, focal heart necrosis, and FH osteomyelitis. However, lesions were not uniform across challenged groups or ages (d 14 and d 21). Alpha diversity of the cecal contents was markedly lower in EC5 and EC11B compared to all treatment groups suggesting that EC exposure during late embryogenesis affect the cecal microbiome up to 21 d posthatch. Additionally, these results highlight the differences in pathogenicity of EC strains isolated from field cases and suggest that hatchery exposure to EC during late embryogenesis is a potential route of introduction into a flock.

The exploration of novel biomarkers to assess poultry health is of paramount importance, not only to enhance our understanding of the pathogenicity of zoonotic agents but also to evaluate the efficacy of novel treatments as alternatives to antibiotics. The present study aimed to investigate potential gut health biomarkers in broiler chicks challenged by Campylobacter jejuni and subjected to a continuous water disinfection program. A total of 144 one-day-old hatched broiler chicks were randomly allocated to four treatment groups with four replicates each, according to the following experimental design: Group A received untreated drinking water; Group B received drinking water treated with 0.01–0.05% v/v Cid 2000™ (hydrogen peroxide, acetic acid and paracetic acid); Group C was challenged by C. jejuni and received untreated drinking water; and Group D was challenged by C. jejuni and received drinking water treated with 0.01–0.05% v/v Cid 2000™. The use of Cid 2000™ started on day 1 and was applied in intervals until the end of the experiment at 36 days, while the C. jejuni challenge was applied on day 18. Potential biomarkers were investigated in serum, feces, intestinal tissue, intestinal content, and liver samples of broilers. Statistical analysis revealed significant increases (p < 0.001) in serum cortisol levels in C. jejuni-challenged broilers. Serum fluorescein isothiocyanate dextran (FITC-d) increased significantly (p = 0.004) in broilers challenged by C. jejuni and treated with drinking water disinfectant, while fecal ovotransferrin concentration also increased significantly (p < 0.001) in broilers that received the drinking water disinfectant alone. The gene expression levels of occludin (p = 0.003) and mucin-2 (p < 0.001) were significantly upregulated in broilers challenged by C. jejuni, while mucin-2 significantly increased in birds that were challenged and received the drinking water disinfectant (p < 0.001). TLR-4 expression levels were significantly (p = 0.013) decreased in both groups that received the drinking water disinfectant, compared to the negative control group. Finally, the C. jejuni challenge significantly increased (p = 0.032) the crypt depth and decreased (p = 0.021) the villus height-to-crypt-depth ratio in the ileum of birds, while the tested disinfectant product increased (p = 0.033) the villus height in the jejunum of birds. Furthermore, the counts of C. jejuni in the ceca of birds (p = 0.01), as well as its translocation rate to the liver of broilers (p = 0.001), were significantly reduced by the addition of the water disinfectant. This research contributes to novel insights into the intricate interplay of water disinfection and/or C. jejuni challenge with potential intestinal biomarkers. In addition, it emphasizes the need for continued research to unveil the underlying mechanisms, expands our understanding of broiler responses to these challenges and identifies breakpoints for further investigations.

ABSTRACT: Clostridial cellulitis or dermatitis affects commercial turkey flocks, primarily as they approach market age. In the field, this disease has been effectively controlled with antibiotics, but alternatives to antibiotics are needed. Bacterin-toxoid vaccination programs have been shown to prevent clostridial diseases in other species, including humans. Results from previous field studies indicate that vaccination with an experimental whole-cell Clostridium septicum (CS) bacterin-toxoid oil emulsion vaccine reduced clostridial dermatitis-associated mortality and antibiotic usage for some commercial turkey flocks, but vaccination was not always efficacious. To improve vaccine efficacy, studies were conducted to optimize the antigenic component of the experimental vaccine and to determine the appropriate antigen to adjuvant ratio, route, and volume for vaccine administration. It was determined that the phase of culture at time of formalin inactivation played a key role in serum antibody titer and larger volume vaccine doses produced higher serum antibody immune response regardless of antigen:adjuvant formulation ratio or route of injection. No significant differences (P > 0.05) were found between formulation ratios or between the subcutaneous and tail head injection sites. Based on these results, we propose to look further into the relationship between culture phase and antigenic components produced by CS under different culture conditions.

A mixed Eimeria spp. challenge model was designed to assess the effects of challenge on broiler chicken performance, intestinal integrity, and the gut microbiome for future use to evaluate alternative strategies for controlling coccidiosis in broiler chickens. The experimental design involved broiler chickens divided into two groups: a control group (uninfected) and a positive control group, infected with Eimeria acervulina (EA), Eimeria maxima (EM), and Eimeria tenella (ET). At day-of-hatch, 240 off-sex male broiler chicks were randomized and allocated to one of two treatment groups. The treatment groups included: (1) Non-challenged (NC, n = 5 replicate pens); and (2) challenged control (PC, n = 7 replicate pens) with 20 chickens/pen. Pen weights were recorded at d0, d16, d31, d42, and d52 to determine average body weight (BW) and (BWG). Feed intake was measured at d16, d31, d42, and d52 to calculate feed conversion ratio (FCR). Four diet phases included a starter d0–16, grower d16–31, finisher d31–42, and withdrawal d42–52 diet. At d18, chickens were orally challenged with 200 EA, 3,000 EM, and 500 ET sporulated oocysts/chicken. At d24 (6-day post-challenge) and d37 (19-day post-challenge), intestinal lesion scores were recorded. Additionally, at d24, FITC-d was used as a biomarker to evaluate intestinal permeability and ileal tissue sections were collected for histopathology and gene expression of tight junction proteins. Ileal and cecal contents were also collected to assess the impact of challenge on the microbiome. BWG and FCR from d16–31 was significantly (p

ABSTRACT: Between 2018 and 2020, over 100 wild turkey fecal samples were collected from the Eastern and Central thirds of the United States, where commercial turkey production is uncommon. We hypothesized that anticoccidial-sensitive Eimeria spp. would be present in wild turkey fecal samples. Samples containing Eimeria spp. oocysts were amplified in vivo. If propagation was successful, the samples were PCR-speciated and subjected to anticoccidial sensitivity testing (AST) for key members of both ionophore and chemical categories of anticoccidial drugs. The purpose of this study was to isolate Eimeria spp. relevant to commercial turkey production that possessed sensitivity to monensin, zoalene, and amprolium. Future research would evaluate the efficacy of wild turkey Eimeria spp. as vaccine candidates for reducing coccidiosis in commercial turkey flocks utilizing single oocyst-derived stocks obtained in the present study.

IntroductionCoccidiosis, caused by parasites of numerous Eimeria species, has long been recognized as an economically significant disease in the chicken industry worldwide. The rise of anti-coccidian resistance has driven a search for other parasite management techniques. Recombinant antigen vaccination presents a highly feasible alternative. Properly identifying antigens that might trigger a potent immune response is one of the major obstacles to creating a viable genetically modified vaccine.MethodsThis study evaluated a reverse immunology approach for the identification of B-cell epitopes. Antisera from rabbits and hens inoculated with whole-sporozoites of E. tenella were used to identify Western blot antigens. The rabbit IgG fraction from the anti-sporozoite serum exhibited the highest reactogenicity; consequently, it was purified and utilized to screen two random Phage-display peptide libraries (12 mer and c7c mer). After three panning rounds, 20 clones from each library were randomly selected, their nucleotide sequences acquired, and their reactivity to anti-sporozoite E. tenella serum assessed. The selected peptide clones inferred amino acid sequences matched numerous E. tenella proteins.Results and ConclusionsThe extracellular domain of the epidermal growth factor-like (EGF-like) repeats, and the thrombospondin type-I (TSP-1) repeats of E. tenella micronemal protein 4 (EtMIC4) matched with the c7c mer selected clones CNTGSPYEC (2/20) and CMSTGLSSC (1/20) respectively. The clone CSISSLTHC that matched with a conserved hypothetical protein of E. tenella was widely selected (3/20). Selected clones from the 12-mer phage display library AGHTTQFNSKTT (7/20), GPNSAFWAGSER (2/20) and HFAYWWNGVRGP (8/20) showed similarities with a cullin homolog, elongation factor-2 and beta-dynein chain a putative E. tenella protein, respectively. Four immunodominant clones were previously selected and used to immunize rabbits. By ELISA and Western blot, all rabbit anti-clone serums detected E. tenella native antigens.DiscussionThus, selected phagotopes contained recombinant E. tenella antigen peptides. Using antibodies against E. tenella sporozoites, this study demonstrated the feasibility of screening Phage-display random peptide libraries for true immunotopes. In addition, this study looked at an approach for finding novel candidates that could be used as an E. tenella recombinant epitope-based vaccine.

IntroductionCoccidiosis caused by the Eimeria spp., an Apicomplexan protozoon, is a major intestinal disease that affects the poultry industry. Although most cases of coccidiosis are subclinical, Eimeria infections impair bird health and decrease overall performance, which can result in compromised welfare and major economic losses. Viable sporulated Eimeria oocysts are required for challenge studies and live coccidiosis vaccines. Potassium dichromate (PDC) is typically used as a preservative for these stocks during storage. Although effective and inexpensive, PDC is also toxic and carcinogenic. Chlorhexidine (CHX) salts may be a possible alternative, as this is a widely used disinfectant with less toxicity and no known carcinogenic associationsMethodsIn vitro testing of CHX gluconate and CHX digluconate exhibited comparable oocyst integrity and viability maintenance with equivalent bacteriostatic and bactericidal activity to PDC. Subsequent use of CHX gluconate or digluconate-preserved Eimeria oocysts, cold-stored at 4°C for 5 months, as the inoculum also resulted in similar oocyst shedding and recovery rates when compared to PDC-preserved oocysts.Results and discussionThese data show that using 0.20% CHX gluconate could be a suitable replacement for PDC. Additionally, autofluorescence was used as a method to evaluate oocyst viability. Administration of artificially aged oocysts exhibiting >99% autofluorescence from each preserved treatment resulted in no oocyst output for CHX salt groups.

Aedes aegypti (Culicidae), the mosquito that causes yellow fever, is regarded as a significant vector for many disease agents. The current study sought to learn more about mosquito mid-gut bacteria and their impacts. Mosquito specimens were gathered in Jeddah city (Saudi Arabia), and their gut bacterial flora was then analyzed. The collected mosquitoes exhibit all the characteristic features of A. aegypti, according to a microscopic study. Utilizing the partial mitochondrial cytochrome oxidase (COI) gene analysis, molecular testing established the identity of A. aegypti as a close relative of previously recognized Aedes species submitted in the GenBank, particularly those from Kenya. Unfortunately, based on the results of RT-PCR, none of the Aedes mosquitoes under study had dengue virus (DENV) present. On nutrient agar plates, the mid-gut bacteria were isolated. A total of 34 Gram-positive bacteria were isolated and identified at the molecular level using the 16S rRNA gene and were divided into the two genera Bacillus and Lysinibacillus within the family Bacillaceae. To our knowledge, this study is the first to examine the normal existence of bacterial flora in the gut of DENV-free mosquitos in Saudi Arabia. It is advised to conduct more research to determine how these bacteria affect the transmission of harmful pathogens carried by mosquitoes. Additionally, further research into the antibacterial and anticancer activities of metabolites extracted from the mid-gut bacteria may help in the development of unique drugs.

IntroductionDrug-sensitive live coccidiosis vaccines have been used to control coccidiosis and renew drug sensitivity in commercial chicken operations. However, only limited species coverage vaccines have been available for commercial turkey producers. This study aimed to assess the effect of an E. meleagrimitis vaccine candidate, with and without amprolium intervention, on performance and oocyst shedding. Additionally, the effect of vaccination, amprolium treatment, and E. meleagrimitis challenge on intestinal integrity and microbiome composition was evaluated.MethodsExperimental groups included: (1) NC (non-vaccinated, non-challenged control); (2) PC (non-vaccinated, challenged control); (3) VX + Amprol (E. meleagrimitis candidate vaccine + amprolium); and 4) VX (E. meleagrimitis candidate vaccine). For VX groups, 50% of the direct poults were orally vaccinated at DOH with 50 sporulated E. meleagrimitis oocysts and were comingled with contact or non-vaccinated poults for the duration of the study. From d10-14, VX + Amprol group received amprolium (0.024%) in the drinking water. All groups except NC were orally challenged with 95K E. meleagrimitis sporulated oocysts/mL/poult at d23. At d29, ileal and cecal contents were collected for 16S rRNA gene-based microbiome analysis.Results and DiscussionVX did not affect performance during the pre-challenge period. At d23-29 (post-challenge), VX groups had significantly (P < 0.05) higher BWG than the PC group. Contacts and directs of VX groups in LS had significantly reduced compared to PC. As anticipated, amprolium treatment markedly reduced fecal and litter OPG for the VX + Amprol group compared to the VX group which did not receive amprolium. The ileal and cecal content results showed that the PC group had different bacterial diversity and structure, including alpha and beta diversity, compared to NC. Linear discriminant analysis Effect Size (LEfSe) identified that Lactobacillus salivarius (ASV2) was enriched in PC’s ileal and cecal content. Compared to NC and PC, the vaccinated groups showed no distinct clusters, but there were similarities in the ileal and cecal communities based on Bray-Curtis and Jaccard distances. In conclusion, these results indicate that vaccination with this strain of E. meleagrimitis, with or without amprolium intervention, caused a very mild infection that induced protective immunity and challenge markedly affected both the ileal and cecal microbiome.

Essential oils (EO) affect performance, intestinal integrity, bone mineralization, and meat quality in broiler chickens subjected to cyclic heat stress (HS). Day-of-hatch Cobb 500 male broiler chicks (n = 475) were randomly divided into four groups. Group 1: No heat stress (Thermoneutral) + control diets with no antibiotics; Group 2: heat stress control + control diets; Group 3: heat stress + control diets supplemented with thymol chemotype (45 ppm) and herbal betaine (150 ppm) formulation EO1; Group 4: heat stress + control diets supplemented with phellandrene (45 ppm) and herbal betaine (150 ppm) formulation EO2. From day 10–42, the heat stress groups were exposed to cyclic HS at 35°C for 12 h (8:00–20:00). BW, BWG, FI, and FCRc were measured at d 0, 10, 28, and 42. Chickens were orally gavaged with FITC-d on days 10 (before heat stress) and 42. Morphometric analysis of duodenum and ileum samples and bone mineralization of tibias were done. Meat quality was assessed on day 43 with ten chickens per pen per treatment. Heat stress reduced BW by day 28 (p < 0.05) compared to thermoneutral chickens. At the end of the trial, chickens that received both formulations of EO1 and EO2 had significantly higher BW than HS control chickens. A similar trend was observed for BWG. FCRc was impaired by EO2 supplementation. There was a significant increase in total mortality in EO2 compared with EO1 EO1 chickens had lower FITC-d concentrations at day 42 than the HS control. In addition, EO1 treatment is not statistically different if compared to EO2 and thermoneutral. Control HS broilers had significantly lower tibia breaking strength and total ash at day 42 than heat-stressed chickens supplemented with EO1 and EO2. Heat stress affected intestinal morphology more than thermoneutral chickens. EO1 and EO2 improved intestinal morphology in heat-stressed chickens. Woody breast and white striping were more common in thermoneutral chickens than heat stress chickens. In conclusion, the EO-containing diet could improve broiler chicken growth during cyclic heat stress, becoming increasingly relevant in antibiotic-free production in harsh climates.

Fenugreek seeds (FSs) are a natural source of bioactive compounds that may modulate the immune system and gut microbiota in broilers. This study examined the effects of dietary fenugreek seed powder on immune-related gene expression and cecal microbiota composition in broilers. A total of 144 broiler chickens were randomly allocated to three dietary groups, CON (0 g/kg FS, FS5 (5 g/kg FS) and FS10 (10 g/kg FS), each with 6 replicates of 8 birds. Ileum tissues and cecal contents were collected on day 42 for the mRNA expression of inflammation and antimicrobial defense-related genes and cecal microbiome diversity, respectively. The results indicated that fenugreek seeds downregulated mRNA-level inflammation and antimicrobial defense-related genes: IL6, IL8L2, CASP6, PTGS2, IRF7, AvBD9, AvBD10, and AvBD11. Moreover, fenugreek seeds altered the cecal microbial community by increasing the population of Firmicutes and decreasing the population of Actinobacteriota, Gemmatimonadota and Verrucomicrobiota at the phylum level and increasing Alistipes, Bacteriodes and Prevotellaceae at the genera level. These findings suggest that fenugreek seeds have a positive impact on the immunological profile and microbiome of broiler chickens, possibly through the interplay of the immune system and the gut microbiome.

Toxoplasma gondii causes a global parasitic disease. Therapeutic options for eradicating toxoplasmosis are limited. In this study, ZnO and Mg-doped ZnO NPs were prepared, and their structural and morphological chrematistics were investigated. The XRD pattern revealed that Mg-doped ZnO NPs have weak crystallinity and a small crystallite size. FTIR and XPS analyses confirmed the integration of Mg ions into the ZnO framework, producing the high-purity Mg-doped ZnO nanocomposite. TEM micrographs determined the particle size of un-doped ZnO in the range of 29 nm, reduced to 23 nm with Mg2+ replacements. ZnO and Mg-doped ZnO NPs significantly decreased the number of brain cysts (p < 0.05) by 29.30% and 35.08%, respectively, compared to the infected untreated group. The administration of ZnO and Mg-doped ZnO NPs revealed a marked histopathological improvement in the brain, liver, and spleen. Furthermore, ZnO and Mg-doped ZnO NPs reduced P53 expression in the cerebral tissue while inducing CD31 expression, which indicated a protective effect against the infection-induced apoptosis and the restoration of balance between free radicals and antioxidant defense activity. In conclusion, the study proved these nanoparticles have antiparasitic, antiapoptotic, and angiogenetic effects. Being nontoxic compounds, these nanoparticles could be promising adjuvants in treating chronic toxoplasmosis.

To assess effects of environmental heat stress (HS) on the local and systemic inflammatory responses to lipopolysaccharide (LPS), broilers were reared under thermoneutral (TN) or cyclic HS conditions. Thermoneutral temperatures followed commercial production settings, with HS broilers exposed to 35 °C for 14 h/day from 4 days onward. At 37 days, HS- and TN-broilers were assigned to either LPS (100 μg/mL) or endotoxin-free phosphate-buffered saline (PBS; vehicle) treatments, eight each to HS- and TN-LPS, four each to HS- and TN-PBS. Treatments were administered by intradermal injection of growing feather (GF) pulps; 10 μL/GF; 12 GF/broiler. Blood and GF were collected before and at 6 and 24 h post-injection to assess leukocyte population changes in GF-pulps and blood, reactive oxygen species (ROS) generation and cytokine expression in GF-pulps, and plasma concentrations of alpha-1 acid glycoprotein (AGP-1). HS-LPS broilers had lower (p ≤ 0.05) infiltration of heterophils and macrophages, ROS generation, and inflammatory cytokine expression in GF-pulps, and lacked the increases in heterophil, monocyte, and plasma AGP-1 concentrations observed in TN-LPS broilers. HS-broilers had similar or greater drops in blood lymphocytes 6 h post-LPS or -PBS injection, respectively, and lower baseline levels (p ≤ 0.05) of circulating T- and B-lymphocytes than TN-broilers. Results indicated that cyclic HS reduced the local and systemic acute inflammatory responses to LPS in broilers, likely impairing their innate defense against microbial infection.

The selection of components within a formulation or for treatment must stop being arbitrary and must be focused on scientific evidence that supports the inclusion of each one. Therefore, the objective of the present study was to obtain a formulation based on ascorbic acid (AA) and Eudragit FS 30D microparticles containing curcumin–boric acid (CUR–BA) considering interaction studies between the active components carried out via Fourier transform infrared spectrometry (FTIR) and differential scanning calorimetry (DSC) to minimize antagonistic effects, and comprehensively and effectively treat turkey poults infected with Salmonella enteritidis (S. enteritidis). The DSC and FTIR studies clearly demonstrated the interactions between AA, BA, and CUR. Consequently, the combination of AA with CUR and/or BA should be avoided, but not CUR and BA. Furthermore, the Eudragit FS 30D microparticles containing CUR–BA (SD CUR–BA MP) showed a limited release of CUR–BA in an acidic medium, but they were released at a pH 6.8–7.0, which reduced the interactions between CUR–BA and AA. Finally, in the S. enteritidis infection model, turkey poults treated with the combination of AA and SD CUR–BA MP presented lower counts of S. enteritidis in cecal tonsils after 10 days of treatment. These results pointed out that the use of an adequate combination of AA and CUR–BA as an integral treatment of S. enteritidis infections could be a viable option to replace the indiscriminate use of antibiotics.

This study compared the minimal inhibition concentrations (MICs) and their effects on the growth kinetics of seven different types of zinc (Zn) compounds and Na2EDTA in the case of three typical commensal beneficial microorganisms (Bacillus subtilis, Lactococcus lactis, and Saccharomyces cerevisiae). The seven Zn compounds included ZnSO4, four Zn–amino acid chelates, and two Zn–EDTA complexes. Both MICs and growth kinetic parameters indicated that different microorganisms show different sensitivities; for example, B. subtilis, L. lactis, and S. cerevisiae were most sensitive to ZnSO4, Na2EDTA, and Zn(NH3)2(Gly)2, respectively. Both ZnEDTA and Zn(NH3)2(Lys)2 improved the growth rate of all beneficial commensal intestinal microorganisms at low concentrations (5–10 mg/L) and showed low toxicity towards all tested strains. At higher concentrations (100–500 mg/L), all compounds decreased the growth rate and increased the lag phase. In conclusion, both growth kinetic parameters and MICs tested effectively measured the inhibitory effects of the test materials; however, growth kinetics provides a more detailed picture of the concentration-dependent effects and those on the mechanisms of microbial growth inhibition.

The primary contaminants in poultry are Salmonella enterica, Campylobacter jejuni, Escherichia coli, and Staphylococcus aureus. Their pathogenicity together with the widespread of these bacteria, contributes to many economic losses and poses a threat to public health. With the increasing prevalence of bacterial pathogens being resistant to most conventional antibiotics, scientists have rekindled interest in using bacteriophages as antimicrobial agents. Bacteriophage treatments have also been investigated as an alternative to antibiotics in the poultry industry. Bacteriophages’ high specificity may allow them only to target a specific bacterial pathogen in the infected animal. However, a tailor-made sophisticated cocktail of different bacteriophages could broaden their antibacterial activity in typical situations with multiple clinical strains infections. Bacteriophages may not only be used in terms of reducing bacterial contamination in animals but also, under industrial conditions, they can be used as safe disinfectants to reduce contamination on food-contact surfaces or poultry carcasses. Nevertheless, bacteriophage therapies have not been developed sufficiently for widespread use. Problems with resistance, safety, specificity, and long-term stability must be addressed in particular. This review highlights the benefits, challenges, and current limitations of bacteriophage applications in the poultry industry.

Small molecules as well as peptide-based therapeutic approaches have attracted global interest due to their lower or no toxicity in nature, and their potential in addressing several health complications including immune diseases, cardiovascular diseases, metabolic disorders, osteoporosis and cancer. This study proposed a peptide, GE18 of subtilisin-like peptidase from the virulence factor of aquatic pathogenic fungus Aphanomyces invadans, which elicits anti-cancer and anti-microbial activities. To understand the potential GE18 peptide-induced biological effects, an in silico analysis, in vitro (L6 cells) and in vivo toxicity assays (using zebrafish embryo), in vitro anti-cancer assays and anti-microbial assays were performed. The outcomes of the in silico analyses demonstrated that the GE18 peptide has potent anti-cancer and anti-microbial activities. GE18 is non-toxic to in vitro non-cancerous cells and in vivo zebrafish larvae. However, the peptide showed significant anti-cancer properties against MCF-7 cells with an IC50 value of 35.34 µM, at 24 h. Besides the anti-proliferative effect on cancer cells, the peptide exposure does promote the ROS concentration, mitochondrial membrane potential and the subsequent upregulation of anti-cancer genes. On the other hand, GE18 elicits significant anti-microbial activity against P. aeruginosa, wherein GE18 significantly inhibits bacterial biofilm formation. Since the peptide has positively charged amino acid residues, it targets the cell membrane, as is evident in the FESEM analysis. Based on these outcomes, it is possible that the GE18 peptide is a significant anti-cancer and anti-microbial molecule.

The use of the USP IV apparatus (flow-through cell) has gained acceptance in recent years due to its versatility and ability to discriminate due to its hydrodynamic conditions. Therefore, the objective of the present study was to develop a discriminative dissolution method in the USP IV apparatus using the open-loop configuration, as well as to propose a method to compare non-cumulative dissolution profiles obtained in the open-loop configuration considering kinetic parameters and validate its predictive power through its comparison with independent and dependent methods using five commercial immediate-release tablet drugs (one reference drug and four generic drugs) of metoprolol tartrate as a model drug. The comparison of the non-accumulated dissolution profiles consisted of determining the geometric ratio of Cmax, AUC0∞, AUC0Cmax, and Tmax (kinetic parameters) of the generic/reference drugs, whereby generic drugs “C” and “D” presented the highest probability of similarity since their 90% confidence intervals were included, or they were very close to the acceptance interval (80.00–125.00%). These results were consistent with the f2, bootstrap f2, and dissolution efficiency approaches (independent models). In conclusion, the proposed comparison method can be an important tool to establish similarity in dissolution profiles and to facilitate the development/selection of new formulations and positively ensure bioequivalence in clinical studies.

Diabetes Mellitus is a metabolic disease that leads to microvascular complications like Diabetic retinopathy (DR), a major cause of blindness worldwide. Current medications for DR are expensive and report multiple side effects; therefore, an alternative medication that alleviates the disease condition is required. An interventional approach targeting the vascular endothelial growth factor (VEGF) remains a treatment strategy for DR. Anti-VEGF medicines are being investigated as the main therapy for managing vision-threatening complications of DR, such as diabetic macular oedema. Therefore, this study investigated the effect of flavonoid naringenin (NG) from citrus fruits on inhibiting early DR in zebrafish. When exposed to 130 mM glucose, the zebrafish larvae developed a hyperglycaemic condition accompanied by oxidative stress, cellular damage, and lipid peroxidation. Similarly, when adult zebrafish were exposed to 4% Glucose, high glucose levels were observed in the ocular region and massive destruction in the retinal membrane. High glucose upregulated the expression of VEGF. In comparison, the co-exposure to NG inhibited oxidative stress and cellular damage and restored the glutathione levels in the ocular region of the zebrafish larvae. NG regressed the glucose levels and cellular damage along with an inhibition of macular degeneration in the retina of adult zebrafish and normalized the overexpression of VEGF as a promising strategy for treating DR. Therefore, intervention of NG could alleviate the domestication of alternative medicine in ophthalmic research.

Abstract The purpose of this pilot study was to evaluate and determine the concentration of prostaglandin GF2α (PGF2α) and isoprostane 8‐iso‐PGF2α in plasma and intestine of specific pathogen-free (SPF) Leghorn chickens challenged with Eimeria maxima, with or without dietary supplementation of curcumin using solid‐phase microextraction and ultra‐performance liquid chromatography/tandem mass spectrometry. Eighty 1-day-old male SPF chickens were randomly allocated to one of four groups with four replicates (n = 5 chickens/replicate). Groups consisted of: (1) Control (no challenge), (2) Curcumin (no challenge), (3) Eimeria maxima (challenge), and (4) Eimeria maxima (challenge) + curcumin. At day 28 of age, all chickens in the challenge groups were orally gavaged with 40,000 sporulated E. maxima oocysts. No significant differences (P > 0.05) were observed in the groups regardless of the treatment or challenge with E. maxima. Enteric levels of both isoprostane 8‐iso‐PGF2α and PGF2α at 7 days and 9 days post-challenge were significantly increased (P

Outbreaks of histomonosis in turkeys are typically initiated by the ingestion of contaminated embryonated eggs of Heterakis gallinarum, potentially present in earthworms and mechanical vectors. Once an outbreak is started, infected turkeys can transmit the disease by horizontal transmission. Factors influencing horizontal transmission of histomonosis are poorly understood. Replication of horizontal transmission in experimental conditions has not been consistent, presenting an obstacle in searching for alternatives to prevent or treat the disease. Two pilot experiments and three validation experiments were conducted in the present study. In pilot experiment 1, one isolate of Histomonas meleagridis (named Buford) was used. Turkeys were fed a low-nutrient density diet corn-soy based (LOW-CS) and raised in floor pens. In pilot experiment 2, another isolate of H. meleagridis was used (named PHL). Turkeys were fed a low-nutrient density diet with the addition of wheat middlings (LOW-WM) and raised in floor pens. In experiment 3, conducted on floor pens, both isolates and diets were used in different groups. In experiment 4, turkeys were raised on battery cages and only the PHL isolate was used. Both diets (LOW-WM and LOW-CS) were used, in addition to a diet surpassing the nutritional needs of young poults (turkey starter, TS). In experiment 5, conducted in battery cages, only the PHL isolate was used, and the LOW-WM and TS diets were in different groups. The horizontal transmission was achieved only with the PHL isolate from all experiments. The transmission rate varied among experimental diets, with the TS diet having the lowest transmission rate in experiments 4 and 5. Variation was observed between experiments and within experimental groups.

Histomonas meleagridis, the etiological agent of histomonosis, is a poultry parasite primarily detrimental to turkeys. Characteristic lesions occur in the liver and ceca, with mortalities in turkey flocks often reaching 80–100%. Chickens and other gallinaceous birds can be susceptible but the disease was primarily considered sub-clinical until recent years. Treating and preventing H. meleagridis infection have become more difficult since 2015, when nitarsone was voluntarily removed from the market, leaving the poultry industry with no approved prophylactics, therapeutics, or vaccines to combat histomonosis. Phytogenic compounds evaluated for chemoprophylaxis of histomonosis have varied results with in vitro and in vivo experiments. Some recent research successes are encouraging for the pursuit of antihistomonal compounds derived from plants. Turkeys and chickens exhibit a level of resistance to re-infection when recovered from H. meleagridis infection, but no commercial vaccines are yet available, despite experimental successes. Safety and stability of live-attenuated isolates have been demonstrated; furthermore, highly efficacious protection has been conferred in experimental settings with administration of these isolates without harming performance. Taken together, these research advancements are encouraging for vaccine development, but further investigation is necessary to evaluate proper administration age, dose, and route. A summary of the published research is provided in this review.

Abstract This study describes a simple method for the large‐scale isolation of pure Toxoplasma gondii tachyzoites and bradyzoites. T. gondii tachyzoites were obtained from infected human foreskin fibroblasts (HFFs) and peritoneal exudates of mice, while tissue cysts containing bradyzoites were collected from chronically infected mice. Harvested cells and brain tissues were incubated in Hanks balanced salt solution (HBSS), containing 0.25% trypsin and 0.5% taurodeoxycholic acid (TDC) for 5 min. Subsequent washes in phosphate buffered saline (PBS) were conducted, and the cell viability of the preparations was good, as determined by flow cytometry and ability to reinfect HFF cells and propagate in mice. The purification procedure allowed for a rapid preparation of pure T. gondii tachyzoites and bradyzoites in sufficient quantity that can be used for downstream procedures. The advantage of the new method is that it is convenient and inexpensive.

Abstract 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 2.0 and P

Tilapia is a widely cultured species native to Africa; these fish are prolific breeders and constitute an economically important fish species supplying higher-quality protein. To meet the global food demand and achieve the UN’s Sustainable Developmental Goals (SDG), the aquaculture industry has conceived of productive solutions with the potential for adaptability, palatability, and profitability. Tilapia may play a vital role with respect to the possibility for sustainability in the nutrition and aquaculture sectors. India contributes to the promotion of aquacultural practices through a structural framework focused on agricultural, environmental, geographical, and socio-economic factors that provide opportunities for tilapia farming. Globally, the Indian aquaculture sector is currently the second largest in terms of aquacultural production but is moving toward different species that meet SDG and facilitate international marketing opportunities. The farming of aquacultural species with innovative technology constitutes an efficient use of resources. Productive research on feeding, disease management, construction, and layout helps overcome the challenges faced in aquaculture. These focused and sustained factors of the aquaculture industry offer a latent contribution to global food security. This review reports on the state of the art, the challenges regarding tilapia aquaculture in India, and the Indian government’s schemes, missions, subsidies, projects and funding related to tilapia production.

The three Bacillus strains present in Norum™ were initially selected by their excellent to good relative enzyme activity (REA) production score for amylase, protease, lipase, phytase, cellulase, β-glucanase, and xylanase. Further studies confirmed that the three isolates also showed an antibacterial activity, Gram-positive and Gram-negative poultry pathogens. Norum™ (Eco-Bio/Euxxis Bioscience LLC) is a Bacillus spore direct-fed microbial (DFM). The Bacillus isolates were screened and selected based on in vitro enzyme production profiles. Moreover, in chickens fed high non-starch polysaccharides, this DFM demonstrated to reduce digesta viscosity, bacterial translocation, increase performance, bone mineralization, and balance the intestinal microbiota. In the present study, we present the whole-genome sequence of each of the three isolates in Norum™, as well as the synergistic, additive, or antagonistic effects on the enzyme production behavior of the three Bacillus strains and their combinations when grown together vs. when grown individually. The whole-genome sequence identified isolate AM1002 as Bacillus subtilis (isolate 1), isolate AM0938 as Bacillus amyloliquefaciens (isolate 2), and isolate JD17 as Bacillus licheniformis (isolate 3). The three Bacillus isolates used in the present study produce different enzymes (xylanase, cellulase, phytase, lipase, protease, and β-glucanase). However, this production was modified when two or more Bacillus strains were combined, suggesting possible synergistic, antagonistic, or additive interactions. The Bliss analysis suggested (p < 0.05) that the combination of Bacillus strains 1–2 and 1–2–3 had intermediate effects and predicted that the combination of Bacillus strains 2–3 could have better effects than the combination of all the three Bacillus strains. In summary, the current study demonstrated the need of selecting Bacillus strains based on quantitative enzyme determination and data analysis to assess the impacts of combinations to avoid antagonistic interactions that could limit treatment efficacy. These results suggest that using Bacillus strains 2–3 together could lead to a new generation of DFMs with effects superior to those already examined in Bacillus strains 1–2–3 and, therefore, a potential alternative to growth-promoting antibiotics. More research utilizing poultry models is being considered to confirm and expand the existing findings.