Comparing the effect of phytobiotic, coccidiostat, toltrazuril, and vaccine on the prevention and treatment of coccidiosis in broilers.

This study compared 2 herbal anticoccidiosis drugs (water-soluble and feed-additive drugs) with monensin coccidiostat, toltrazuril (TTZ, anticoccidiosis drug), and Livacox Q (anticoccidiosis vaccine) in terms of their effects on the prevention and treatment of coccidiosis in broilers. In this study, 280 Ross 308 broiler chickens (a mix of both genders) were used in a completely randomized design with 7 treatments and 5 replications each including 8 chickens per replicate. On d 21 of rearing, all experimental groups, except for the negative control group (NC), were challenged with a mixed suspension of common strains of Eimeria, and the intended indices were assessed, including performance indices, number of oocysts per gram (OPG) of feces, intestinal injuries, and the total number of intestinal bacteria. In addition, the NC and the group receiving the monensin had greater body weight gain (BWG) (P < 0.05). At the end of week 6, the monensin group had the highest feed intake (FI), while the water soluble medicine treatment resulted in the lowest feed intake (P < 0.05). Regarding the lesion scores on day 28, the highest and lowest rates of jejunal injuries were observed in the positive control group (PC), the monensin and vaccine group respectively. The rate of oocysts excretion (oocysts per gram of feces = OPG) on different days was higher in the PC group, and the use of monensin could further reduce excretion compared to the other groups (P > 0.05). Based on a comparison of the population of lactic acid bacteria between the NC and both medicinal plant treated groups, the use of these products could increase the population of these types of bacteria. Moreover, the population of Escherichia coli was less considerable in the NC and herbal powder groups (P < 0.05). Overall, similar to commercial medicines, the herbal medicines used in this project can be effective in the prevention and treatment of coccidiosis and can improve profitability in broiler rearing centers by improving intestinal health.

The effect of peroxisome proliferator-activated receptor gamma (PPARγ) as a mediator of dietary fatty acids and thiazolidinedione in pulmonary arterial hypertension induced by cold stress of broilers.

The objective of this experiment was to evaluate the effects of dietary fish oil and pioglitazone as peroxisome proliferators-activated receptor gamma (PPARγ) activating ligands on the reduction of cold-induced ascites in broiler chickens. A total of 480 one-day-old (Ross 308) male chicks were randomly allocated to four treatment groups with eight replicates of 15 birds each. The following treatments were used: 1) ambient temperature (negative control), with basal diet; 2) cold-induced ascites (positive control), with basal diet; 3) cold-induced ascites, with basal diet +10 mg/kg/day pioglitazone and 4) cold-induced ascites, with basal diet +1% of fish oil. When compared with the positive control, body weight gain was higher (P ≤ 0.05) for broilers fed diets containing fish oil and pioglitazone at 28, 42, and 0-42 d. Broilers under cold-induced ascites had the highest blood pressure at 21 and 42 d, while fish oil and pioglitazone treatment reduced the blood pressure (P ≤ 0.05). Red blood cells, white blood cells, hematocrit, erythrocyte osmotic fragility, bursa of Fabricius and spleen weights were improved (P ≤ 0.05) for chickens fed fish oil diets and pioglitazone compared to the cold-induced ascites (positive control). Exposure to cold temperature resulted in an increase in plasma T3 and T3/T4 ratio and decline in plasma T4 (P ≤ 0.05). In conclusion, PPARγ agonist pioglitazone and fish oil as source of omega-3 polyunsaturated fatty acid could be used as a strategy to reduce the negative effects of pulmonary arterial hypertension and ascites in broiler chickens.

Peroxisome proliferator-activated receptor gamma (PPARγ) activation: a potential treatment for ascites syndrome in broiler chickens.

Ascites (serous fluid accumulation in the abdominal cavity) has been observed worldwide in fast growing broilers. Pulmonary vascular remodeling is an important pathological feature of broiler ascites syndrome. Peroxisome proliferators-activated receptor gamma (PPARγ) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) are expressed in pulmonary vascular endothelial cells and vascular smooth muscle cells (VSMC) where they participate in the regulation of normal pulmonary vascular function. The objective of the present study was to investigate the effects of omega-3 fatty acids (found in fish oil) and pioglitazone (PIO) as natural and synthetic PPARγ ligands supplementation on PPARγ and PGC-1α expression in the prevention of pulmonary arterial hypertension (PAH) syndrome in broiler chickens. The experiment was conducted with 4 treatment groups: 1) negative control, normal temperature conditions with basal diet; 2) positive control, low-temperature conditions with basal diet; 3) positive control + 10 mg PIO/kg of weight/d and 4) positive control + 1% FO. Each treatment had 5 replicates. Ascites heart index (RV/TV) was significantly (P < 0.05) reduced in chickens receiving FO (0.20) and PIO (0.21) compared to the positive control group (0.26). The addition of PIO in broilers under cold-induced ascites significantly increased the expression of PPARγ (9.44) and PGC-1α (5.81) genes in lung tissue compared to the negative control group (1.03, P < 0.05). Proliferative indexes of VSMC in pulmonary arteries such as PMT, PIT, and percentage wall thickness were significantly elevated in positive control group, indicating that pulmonary vascular remodeling occurred following VSMC proliferation in ascites. The vessel internal diameter was increased in FO and PIO groups. Based on these results, activation and expression of PPARγ and PGC-1α genes as a critical regulator of pulmonary artery smooth muscle cell using ligands, especially PIO, can be effective in reducing the incidence of PAH in broiler chickens.

The Effects of Glycerophospholipid Nanomicelles on the Cryotolerance of Frozen-Thawed Rooster Sperm.

Semen banking is an efficient method of artificial insemination for commercial breeders. However, the cryopreservation process induces severe damages to plasma membranes, which leads to reduced fertility potential of thawed sperm. The replacement of membrane lipids with oxidized membrane lipids repairs the cell membrane and improves its stability. The aim of this study was to investigate the effects of glycerophospholipid (GPL) nanomicelles on the cryosurvival of thawed rooster semen. Semen samples were collected from six 29-week Ross broiler breeder roosters, then mixed and divided into five equal parts. The samples were diluted with the Beltsville extender containing different concentrations of GPL according to the following groups: 0 (GPL-0), 0.1% (GPL-0.1), 0.5% (GPL-0.5), 1% (GPL-1), and 1.5% (GPL-1.5), then diluted semen was gradually cooled to 4°C during 3 hours and stored in liquid nitrogen. The optimum concentration of GPL was determined based on the quality parameters of thawed sperm. Our results showed sperm exposed to GPL-1 had significantly increased motion parameters and mitochondrial activity. The percentages of viability and membrane integrity were significantly higher in the GPL-1, and GPL-1.5 groups compared with the other groups (p < 0.05). Moreover, the lowest rate of apoptosis and lipid peroxidation were observed in the GPL-1 and GPL-1.5 groups in comparison with the frozen control group. Our findings indicated that membrane lipid replacement with GPL nanomicelles (1% and 1.5%) could substitute for damaged lipids in membranes and protect sperm cells against cryoinjury.

Research Note: Evaluation of two methods for adding cryoprotectant to semen and effects of bovine serum albumin on quality characteristics of cryopreserved rooster spermatozoa.

Chicken semen cryopreservation is a tool for programs of genetic diversity management and endangered breeds conservation. Due to physiological features, the fertility rates of cryopreserved poultry sperm are lower than mammal species. Thus, improvement of the semen cryopreservation methods is required. A first study was performed by a 2 × 2 factorial design consisting of 2 methods of adding the cryoprotectant [Direct or Diluted (mixed with extender medium)] and 2 cryoprotectants (glycerol and dimethylacetamide). Then sperm quality indicators were evaluated after freezing. A second study with a 2 × 2 design was conducted to evaluate the effectiveness of bovine serum albumin (BSA) on the optimization of 2 different extenders (Lake and Animal Sciences Group [ASG]). Viability and motility variables were evaluated before and after freezing. There was no significant difference in sperm viability and motility variables between Direct or Diluted methods. Supplementation of extenders with BSA improved most of the sperm motility variables in both extenders before and after freezing. Progressive sperm, non-progressive sperm before freezing, and all post-thaw sperm motility parameters, except amplitude of lateral head displacement and beat-cross frequency, were increased in BSA-supplemented extenders (P < 0.05), and BSA improved sperm viability in ASG extender after thawing (P < 0.05). After thawing, the interaction between extender and BSA (P < 0.05), eliminated the differences between the 2 BSA-supplemented media in curvilinear velocity, straight-line velocity, average path velocity, and amplitude of lateral head displacement which were higher in non-supplemented ASG extender than nonsupplemented Lake medium. In conclusion, the direct or diluted methods of adding glycerol or dimethylacetamide, did not significantly affect the post-thaw sperm characteristics. BSA positively affected most of the post-thaw sperm motility indicators regardless of the type of extender and resulted in significantly higher post-thaw sperm viability in ASG medium.

The effect of methyl-beta-cyclodextrin on DNA absorption and quality of posttransfected sperm.

Sperm can be selected as a natural vector for the production of transgenic animals. Methyl-beta-cyclodextrin (MBCD) removes cholesterol from the phospholipid membrane of sperm and improves the efficiency of DNA uptake by sperm. In experiment 1, fresh sperm was treated with various concentrations of MBCD. The direct effects of MBCD on sperm parameters were monitored. In experiment 2, different concentrations of MBCD (0, 1, 2, and 4 mmol) were assessed for the transfection of genetically exogenous construction to rooster sperm. Washed semen was divided into 5 equal groups for the incubation and transfection with a pcDNA3.1+/hG-CSF vector (exogenous DNA) as follows; Treatment I-Control (washed semen without DNA); Treatment II-Control (washed semen with DNA); Treatment III-(washed semen incubated with DNA and 1 mmol MBCD); Treatment IV-(washed semen incubated with DNA and 2 mmol MBCD); and Treatment V-(washed semen incubated with DNA and 4 mmol MBCD). We demonstrated that rooster spermatozoa spontaneously can uptake exogenous DNA; this was assessed using exogenous DNA amplification (sperm genomic DNA used as a template for PCR reaction) after DNase I treatment. In addition, total motility (TM), progressive motility (PM), velocity parameters [curvilinear velocity (VCL), straight linear velocity (VSL), sperm track straightness (STR), linearity (LIN)], membrane integrity (MI), and membrane functionality were posttransfectionally evaluated. The concentrations of 1 and 2 mmol MBCD significantly (P < 0.05) improved the motion characteristics and membrane integrity of fresh sperm. The presence of hG-CSF in rooster sperm was detected by PCR and based on sperm analyses MBCD (1 mmol) improved the percentage of motility (98.9 ± 0.81), membrane functionality (64 ± 1.64), and MI (76.2 ± 1.65) after transfection when compared with the other groups (P < 0.05). For the production of transgenic chicken, hens were inseminated (AI) by transfected sperm treated with 1 and 0 mmol MBCD. A PCR analysis of the blood samples and dead embryo tissues of chicks did not reveal the transgene integration.

Effects of dietary alpha-lipoic acid supplementation on the seminal parameters and fertility potential in aging broiler breeder roosters.

High levels of polyunsaturated fatty acids in avian sperm cause more susceptibility to lipid peroxidation. Aging in roosters reduces the antioxidant capacity of sperm and thus fertility. The purpose of this study was to investigate the effects of different levels of alpha-lipoic acid (ALA) as a feed supplement to improve the semen quality and fertility parameters of aged broiler breeder roosters and identification of its most effective level. A total of forty-two roosters at 45 wk of age were randomly assigned to 7 treatments (0, 15, 40, 70, 95, 120, and 145 mg ALA/bird per day) for 8 wk. Semen parameters and body weight were assessed biweekly, and testosterone plasma levels were determined in the 8th wk of the experimental period. Artificial insemination was performed at the end of the experiment to evaluate the fertility potential. The dietary administration of ALA had no significant effects on body weight, semen volume, average path velocity, linearity, straightness, wobble, the amplitude of lateral head displacement, beat-cross frequency, sperm concentration, morphology, plasma testosterone level, fertility, or hatchability (P > 0.05). Alpha-lipoic acid supplementations resulted in a significant decrease in seminal malondialdehyde concentration and immotile (type D) sperms (P < 0.05). The total motility, progressive motility (types A + type B sperms), curvilinear velocity, straight-line velocity, viability, and membrane integrity of sperm improved with ALA dietary supplementations (P < 0.05). With increasing ALA levels, improvement in semen parameters had an incremental trend until the level of 95 mg ALA. Thus, 95 mg dietary ALA as an antioxidant supplement can improve semen quality of aging breeder roosters while higher doses resulted in no further improvement.

Sodium bisulfate feed additive aids broilers in growth and intestinal health during a coccidiosis challenge.

Sodium bisulfate (SB) was evaluated on its ability to improve broiler growth and intestinal structure with(out) a coccidia challenge. One thousand two hundred Cobb500 day-old males were randomly assigned within 4 experimental groups with a 2 × 2 factorial design, with (out) SB in the diet and with(out) a day 0 coccidia challenge using a 10× dose of a commercial vaccine. At day 7, oocysts per gram of feces were determined. At day 0, 14, 28, and 41, BW and feed consumption were measured. At day 21, 20 birds per treatment were subjectively scored for coccidia lesions, and jejunal histologic samples were collected for villi measurements. Twenty additional birds were given fluorescein isothiocyanate-dextran to determine gut permeability. At day 41, 10 birds per treatment had histologic samples collected. Statistical analysis was conducted in JMP Pro 14 using GLM procedure to compare disease state and diet. Means were separated using Dunnett's test (P ≤ 0.05) with the nonchallenged standard diet treatment that is considered the control. All parameters measured indicated an effect due to the coccidia inoculation. Therefore, effects of diet on (non)challenged treatments were determined using a Student t test (P ≤ 0.05). Limited differences due to diet were seen for the nonchallenged production data. Sodium bisulfate had a thinner villi base width (P = 0.04) on day 21 and greater villi height (P = 0.03), smaller base width (P = 0.04), thicker muscularis (P = 0.03), and lower crypt: height ratio (P = 0.01) on day 41. Challenged SB had similar gut permeability to the nonchallenged control (P = 0.94) on day 21. There was no difference in flock uniformity, feed intake, oocysts per gram of feces, or lesion scores between challenged treatments. Challenged SB had greater BW on day 14 (P < 0.0001), 28 (P < 0.0001), and 41 (P = 0.02). Feed conversion ratio from day 0 to 14 was also lower (P = 0.0002). Challenged SB had smaller crypts (P = 0.02) and therefore a smaller crypt: height ratio (P = 0.03) on day 21. Challenged control had a larger apical width (P = 0.03) and thicker muscularis (P = 0.04) on day 41. Overall, the addition of SB during coccidial enteropathy aided in BW, feed conversion ratio, and villi health with no observed effects on parasite cycling.

The effect of sodium bisulfate and coccidiostat on intestinal lesions and growth performance of Eimeria spp.-challenged broilers.

Coccidiosis is a high-prevalence disease that annually entails huge costs for the poultry industry. Control of coccidiosis in poultry production is based on the use of coccidiostats and vaccines. However, along with the problem of drug resistance, there is a concern about food safety and drug residues in poultry products. The objective of this study was to evaluate the effect of sodium bisulfate (SBS) in comparison with monensin (M) and their combination (SBSM) effects on controlling coccidiosis in broilers. In a randomized design, 300 chickens (Ross 308) were divided into 5 treatments and 4 replications (15 birds per replicate). All birds, except the negative control (NC), were orally inoculated with 4 Eimeria species on 14 D of age. Treatments included were as follows: NC, an unsupplemented basal diet, nonchallenged; positive control, a basal diet unsupplemented, challenged with Eimeria spp; a basal diet supplemented with 5 g/kg of SBS; a basal diet supplemented with 1 g/kg of M; and a basal diet supplemented with 5 g/kg SBS and 1 g/kg M (SBSM). Oocyst shedding per gram (OPG) of the faecal sample from each experimental unit was counted on 5 to 14 D after inoculation. Two chicks from each experimental unit were euthanized to investigate intestinal lesions on day 5 after inoculation. The NC birds showed the highest BW gain and the lowest feed conversion ratio. The birds in the SBSM group had improved feed consumption compared with the M group in the prechallenge period (P < 0.05). All supplemented treatments resulted in a significant decrease in OPG. The M and SBSM treatments showed more efficacy than the SBS group (P < 0.05) in reducing OPG. There was a significant reduction in cecal lesions owing to supplementation with SBS, but the effect of SBS in the upper part of the intestine was lower than the M and SBSM groups (P < 0.05). Based on the results of this study, SBS has protective effects against coccidiosis in ceca, and the combination of M and SBS (SBSM) did not show any further improvement effect compared with M alone on the control of coccidiosis.

The efficiency of synthetic polymers to ameliorate the adverse effects of Aflatoxin on plasma biochemistry, immune responses, and hepatic genes expression in ducklings.

To evaluate the effect of molecularly imprinted polymers as a synthetic polymer (TMU95) and commercial toxin binder (CTB) on aflatoxins (AFs) toxic effects on hepatic gene expression, and the biochemical and immunological parameters in ducklings, 240 four-day-old ducklings were randomly allocated into six groups with four replicates of 10 ducklings per each. The experimental groups were as follows: Negative control (basal diet without any additive or AFs), Negative control + TMU95 (5 g/kg feed), Negative control + CTB (Zarinbinder, Vivan Group, Mashhad, Iran. 5 g/kg feed), Positive control (0.2 mg AFs/kg feed), Positive control + TMU95 (5 g/kg feed), and Positive control + CTB (5 g/kg feed). On day 14, livers were collected (8 per treatment) to evaluate change in the expression of genes involved in AFs biotransformation (cytochrome P450 1A1 and 2H1) and antioxidant function (glutathione S-transferase). Several biochemical biomarkers and immune responses were also recorded. Compared with the negative control group AFs treatment significantly decreased plasma total cholesterol, triglyceride and increased the aspartate-aminotransferase (AST), alanine-aminotransferase (ALT) and alkaline phosphatase (ALP) activity (P ≤ 0.01). Cellular immune responses to the phytohemagglutinin-and 2, 4-dinitro 1-chlorobenzene skin test were significantly influenced by dietary aflatoxins (P ≤ 0.01) but a humoral immune response to Newcastle disease virus/vaccine was not affected (P ≥ 0.01). Compared with negative control group, the genes associated with AFs biotransformation were downregulated, whereas the gene associated with the antioxidant function was upregulated in birds fed AFs. The CTB supplement in contaminated feed could alleviate AFs adverse effects on cellular immunity, ALT concentration, and cytochrome P450 2H1 gene expression partially, whereas TMU95 could not ameliorate the adverse effects of AFs on the traits studied, except for ALP. The data suggest that TMU95 may alleviate some of the toxic effects of aflatoxins in duckling and it might prove to be beneficial in the reduction of aflatoxicosis adverse effect in poultry when used in combination with other aflatoxin management practices.

The effect of capsulated and noncapsulated egg-yolk-specific antibody to reduce colonization in the intestine of Salmonella enterica ssp. enterica serovar Infantis-challenged broiler chickens.

The antibacterial properties of egg yolk antibodies have been known for many years. Enhanced antibiotic resistance has resulted in increased need for using these antibodies as an alternative. In the present study, generation, capsulation, and inhibition growth properties of IgY directed against Salmonella enterica subsp. enterica serovar Infantis (SI) were evaluated. White Leghorn layer hens were immunized using whole cell of inactivated SI. Salmonella Infantis-specific antibody activities in sera and egg yolk were determined by ELISA. A total of 480 one-day-old male "Cobb 500" chicks were randomly divided into 8 groups, with 6 replications of 10 birds kept for 21 D. All birds from 7 challenged groups were orally inoculated with 1 mL of SI suspension (1 × 107 CFU/mL) at 3 and 4 D of age. Two groups were dietary supplemented with 5 g/kg immune powdered yolk or nonimmune powdered yolk. One group was dietary supplemented with 12.8 g/kg capsulated immune yolk (CIY). Two groups were given 8.3 mL/L of immune water-soluble yolk or nonimmune water-soluble yolk fraction in drinking water. In the antibiotic group, 1 mL/L Enrofloxacin 10% was added to drinking water. All supplements except for the antibiotic (on Day 4 for 10 D) were added on day one and continued during the experiment. Negative and positive control groups received no supplements. During the experiment, among the challenged groups, the minimum SI cecal colonization and the lowest isolation of SI from the liver (P < 0.01) was observed in the antibiotic group. Following antibiotic group, in the group receiving CIY, colonization of bacteria in ceca and liver was significantly reduced during the second and third weeks of the experiment (P < 0.01). According to the results, capsulated specific IgY has a beneficial effect in reducing the colonization of Salmonella under the conditions of this study in comparison with other forms of IgY antibody.

The effectiveness of a dietary direct-fed microbial and mannan oligosaccharide on ultrastructural changes of intestinal mucosa of turkey poults infected with Salmonella and Campylobacter.

Salmonella and Campylobacter are considered major public health burdens worldwide, and poultry are known to be one of the main reservoirs for these zoonotic pathogens. This study was conducted to evaluate the effect of a commercial probiotic or direct-fed microbial (DFM) Calsporin (CSP), and prebiotic or mannan oligosaccharide (MOS) (IMW50) on ultrastructural changes and the villous integrity of intestinal mucosa in turkey poults challenged with Salmonella and Campylobacter. A 21-day battery cage study was conducted using 4 dietary treatments including a basal diet (corn and soybean-based) nonsupplemented and uninfected as a negative control (NC); basal diet supplemented with 0.05% DFM (CSP); basal diet supplemented with 0.05% MOS (IMW50); and basal diet supplemented with 0.05% mixture of DFM and MOS at equal proportions. Female large white turkey poults aged 336 days were obtained from a local commercial hatchery and randomly distributed in electrically heated battery cages with 12 treatments of 4 replicates per treatment containing 7 poults per pen. The first 16 pens were not infected with bacteria, poults in pens 17-32 were orally challenged at day 7 with 105 cfu Salmonella Heidelberg, and the poults in pens 33-48 were orally challenged at day 7 with 105 cfu Campylobacter jejuni. Feed and water were provided ad libitum throughout the study. At day 21, ileal tissue samples from 1 bird per cage were collected for intestinal integrity and ultrastructural examination by scanning and electron microscopy. DFM and MOS supplementation was effective in both challenged and nonchallenged (not infected with Salmonella and Campylobacter) birds. Goblet cells and mucus were increased, with the presence of large numbers of segmented filamentous bacteria in DFM- and MOS-supplemented groups compared with birds in control treatments. The number and size of villi were reduced in poults exposed to Salmonella and Campylobacter. Results show that CSP and IMW50 provide protection of ileal mucosal integrity in poults exposed to Salmonella or Campylobacter.

Plant extract supplementation as a strategy for substituting dietary antibiotics in broiler chickens exposed to low ambient temperature.

The present study was conducted to investigate the effects of two plant extracts as alternatives to dietary antibiotics in broiler chickens exposed to low ambient temperature. A total of 300 one-day-old male broiler chickens were randomly assigned to four dietary treatments (5 replicate pens; 15 broiler chickens each) which consisted of starter (d 0 to 10), and grower (d 10 to 28) diets. Dietary treatments included a basal diet (negative control, NC) and three similar diets that were either supplemented with 200 mg/kg of Prosopis farcta extract (PFE), Rhus coriaria L. extract (RCE) or an antibiotic premix containing oxytetracycline (positive control, PC). In order to simulate low ambient temperature, room temperature was maintained at 32°C during the first 3 d of the trial and afterwards, the temperature was gradually reduced by approximately 1.5°C each day to 14°C on d 21. PFE and PC treatments exerted a significant effect on body weight gain at d 28. Diet PFE was effective in reducing mortality when compared with diet NC (p < 0.05). Furthermore, diet PFE caused increases in ileal  digestibility of gross energy, dry matter and organic matter when compared with diet NC (p < 0.05). Diets PFE and PC decreased coliforms, total aerobic bacteria and total anaerobic bacteria loads in the caeca when compared with diet NC (p < 0.05). Moreover, the addition of PFE to the diet improved villous height in all small intestinal segments as well as villous height:crypt depth ratio in the duodenum when compared with diet NC (p < 0.05). The results indicated that PFE is not only a valid alternative to oxytetracycline under cold stress conditions, with no antibiotic resistance, but also has the potential to increase the resistance of broiler chickens against ascites syndrome. Moreover, the addition of RCE at the concentration of 200 mg/kg to the diet was not sufficient to improve the performance of broiler chickens (similar to diet PC) but maybe more effective at higher concentrations.

Effect of a direct-fed microbial and prebiotic on performance and intestinal histomorophology of turkey poults challenged with Salmonella and Campylobacter.

Salmonella and Campylobacter are leading human foodborne pathogens commonly associated with poultry and poultry products, and several methods to control these pathogens have been applied to poultry production. This study was conducted to evaluate the effect of CALSPORIN, (CSP), a direct-fed microbial (DFM), and yeast cell wall (Saccharomyces cervisiae, IMW50, a mannanoligosaccharide (MOS)-based prebiotic, on performance, levels of Salmonella and Campylobacter in the feces, and intestinal histomorphometry in turkey poults. A 21-day battery cage study was conducted using 4 dietary treatments, including: an unsupplemented basal diet (corn and soybean-based) as negative control (NC); basal diet supplemented with 0.05% DFM; basal diet supplemented with 0.05% MOS; and basal diet supplemented with 0.05% mixture of DFM and MOS at equal proportions. Female Large White turkey poults (n = 336) were randomly distributed in 6 electrically-heated battery cages with 4 treatments and 12 replicates per treatment (7 poults per replicate pen). The first 16 pens were not inoculated with bacteria, while poults in pens 17 to 32 were orally challenged at day 7 with 105 CFU Salmonella Heidelberg and the poults in pens 33 to 48 were orally challenged at day 7 with 105 CFU Campylobacter jejuni. Feed consumption, body weight, and feed conversion ratio were measured weekly and at the end of the experiment. At day 21, fresh fecal samples from each pen were collected for Salmonella and Campylobacter enumeration and ileal tissue samples were collected from 1 bird per pen for histomorphology examination. DFM and MOS supplementation was accompanied with reduced levels of Salmonella shed by the treated birds compared to the control group, and with increased body weight (P ≤ 0.05). The surface area of villi increased in the MOS-supplemented group compared to the control group (P ≤ 0.05). There was a significant difference in V:C ratio between supplemented groups and control group (P ≤ 0.05). Based on these results, there is potential for CALSPORIN and IMW50 to reduce Salmonella shedding in feces, enhance ileal mucosal health, and improve growth performance of turkey poults.

Effect of serum anti-phage activity on colibacillosis control by repeated phage therapy in broilers.

Anti-phage activity of serum is of importance in repeated phage therapy. Higher serum anti-phage activity has been associated with greater susceptibility of phages to neutralisation and phage therapy failure. In this study, in vivo and in vitro survivability and immunogenicity of four coliphages (TM1, TM2, TM3 and TM4) were investigated in naive chickens and chickens pre-immunised with phage TM1. Furthermore, two phages that displayed different survivability and immunogenicity (TM1 and TM3) were compared with respect to their efficacy in treating naive or pre-immunised (TM1) chickens suffering from colibacillosis. The efficacy of the treatments was evaluated based on body weight, relative organ weights, mortality, E. coli counts in the lungs as well as severity and frequency of internal organ lesions. At the end of the experiment, both naive and pre-immunised chickens treated with TM3 showed significantly lower mortality and higher body weights than untreated chickens and those treated with TM1. The same trend was observed in incidence and severity of organ lesions as well as relative spleen weight. However, naive chickens treated with TM1 also showed a shortened inflammation period as indicated by spleen weights. E. coli counts in the lungs of chicken treated with TM3 were lower than those of chickens treated with TM1 on days 3 and 10 post challenge. These data indicate that the outcome of phage therapy and the impact of serum anti-phage activity are highly phage-type dependent in broilers.

Comparative evaluation of aflatoxin and mineral binding activity of molecular imprinted polymer designed for dummy template using in vitro and in vivo models.

The amelioration of aflatoxicosis in ducklings was examined by feeding molecular imprinted polymer (MIP) synthesized to target the aflatoxin B1 (AFB1) analog molecule [5, 7-dimethoxycoumarin (DMC)] as a smart and novel toxin binder in comparison to a commercial toxin binder (CTB). MIP was characterized with some techniques, such as scanning electron microscopy (SEM) and thermal gravimetric analysis (TGA) that showed morphology and thermal resistance of MIP, respectively. The SEM showed that more pores and big cavities were formed in the network of the MIP. TGA plots of MIP showed dehydration at temperatures of about 31 °C-200 °C and gradual decomposition until 300 °C. The in vitro and in vivo ability of MIP and CTB to adsorb some minerals (Ca, Cu, Mg, P and Zn) was evaluated. The in vivo efficacy of MIP as an aflatoxins (AFs) binder in duckling exposed to aflatoxin-contaminated feed from 4 to 18 d of age was also compared to that of the CTB. A total of 240 4-d-old ducklings were assigned to experimental diets. Each diet was replicated four times with 10 birds in each pen, and the initial body weight classes were considered as blocks. The experimental treatment diets included: 1- Control (basal diet without any additive or AFs), 2- MIP (5 g/kg diet), 3- CTB (5 g/kg diet), 4- AFs (0.2 mg/kg diet), 5- AFs + MIP (0.2 mg/kg + 5 g/kg diet), and 6- AFs + CTB (0.2 mg/kg + 5 g/kg diet). In vitro study may clearly reflect the possibility of adsorbing minerals by CTB as compared to MIP whilst the results were not confirmed by in vivo study and it seems that reduction in plasma levels of minerals is related to the adverse effects of AFs. Livers showed histopathological alterations, with bile-duct proliferation of all aflatoxin fed ducklings. Macroscopic study indicated liver of birds fed diets containing AFs showed abnormal signs including yellowish, friable and rounded shape without protective effect of MIP and CTB. The results obtained from in vivo study indicate the partial alleviation of the adverse effects of AFs in ducklings by dietary supplementation of MIP and CTB (5 g/kg) on plasma albumin, Zn, Mg and Ca.

The first in vivo application of synthetic polymers based on methacrylic acid as an aflatoxin sorbent in an animal model.

This study attempts to evaluate the potential aflatoxin binder activity of a molecularly imprinted polymer (TMU95) synthesized to target the aflatoxin B1 (AFB1) analog molecule in comparison to a commercial toxin binder (CTB). Adsorption experiments were carried out to assess the ability to bind to AFB1 at various pH values. The strength of binding was investigated by the chemisorption index. The isothermal analysis was used to determine the maximum adsorption capacity values. The ability of TMU95 and CTB to adsorb essential minerals was evaluated and the obtained data suggested that CTB would significantly reduce availability of them compared to TMU95. The in vivo efficacy of TMU95 as an aflatoxin (AF) binder in duckling exposed to aflatoxin-contaminated feed from 4 to 18 days of age in comparison to the CTB was also assessed. TMU95 and CTB were effective in reducing the adverse effects caused by AFs on feed conversion ratio of duckling (p ≤ 0.01), and also showed a minor reduction of injuries caused by AFs on visceral organs enlargement (p ≤ 0.01). It was concluded that TMU95 could absorb AFB1 in vitro efficiently and had beneficial health effects that could alleviate some of the toxic effects of AFs on growing duckling performance similar to CTB.

Synergistic effect of phage therapy using a cocktail rather than a single phage in the control of severe colibacillosis in quails.

Infections associated with avian pathogenic Escherichia coli (APEC) cause severe economic losses to the poultry industry. The study presented herein investigated the in vivo performance of a single phage with prolonged in vivo and in vitro survivability alone or in combination with 3 other selected phages in treating colibacillosis in quails. Japanese quails (N = 360) were randomly assigned to 6 treatment groups with 4 replicate pens. Birds from the control groups (groups I, II, and III) were treated with 200 µL sterile PBS (pH 7.4), 200 µL of the selected phage (1010 pfu; TM3) or a cocktail of 4 phages (TM3 plus TM1, TM2, and TM4), respectively. Groups IV, V, and VI were challenged with 200 µL E. coli (108 cfu; O78:K80 and O2:K1) and treated with i.m. injection of 200 µL sterile PBS, phage TM3, or cocktail of 4 phages, respectively. Based on the results of the present study, the total mortality rate decreased from 46.6% in the untreated E. coli-challenged group to 26.5% and 13.6% in the E. coli-challenged group treated with single phage or phage cocktail, respectively. The body weights of birds treated with the phage cocktail were higher than the body weights of untreated birds on days 7, 14, and 21 post-challenge (P < 0.05). In addition, total viable cell counts of E. coli in the lungs of birds treated with the phage cocktail were lower than those of birds treated with phage TM3 on days 3 and 10 post-challenge (P < 0.05). Moreover, the incidence and severity of lesions in lungs, heart, and liver were found to be significantly less in the E. coli- challenged group treated with the phage cocktail. In conclusion, this study indicates that a phage cocktail may be more efficient in treating colibacillosis than a single phage possibly due to a synergistic effect between the individual phages.

In vitro improvement of quail primordial germ cell expansion through activation of TGF-beta signaling pathway.

Avian primordial germ cells (PGCs) have valuable potentials to cell-based approaches for transgenic bird production. In this regard, improvement of avian PGC expansion in vitro is necessary. Among experimental avian species, quail is a good model for transgenic technology, especially due to its short generation time. In the present study, we have examined the proliferative effects of transforming growth factor ß (TGF-ß) on the quail PGCs. After isolation of quail PGCs from blood (Hamburger-Hamilton [HH stages 13-15]) and gonads (HH stages 28-30), these cells were cultured on quail embryonic fibroblasts (QEF). Our results indicated th at cultured gonadal-derived PGCs proliferated 400 times in comparison to 100 times for blood PGCs over 40-50 days. Upon in vitro exposure to TGF-ß inducers by Activin or the inducer of definitive endoderm 1 (IDE1) small molecule, the number of gonad PGCs significantly increased to 26% and 64%, respectively. In contrast, inhibition of the TGF-ß signaling pathway by SB431542 resulted in a significant reduction in the numbers of PGCs (P < 0.001). Moreover, Phosphorylation of SMAD2/3 in the IDE1 group was higher compared to the Activin-treated ones. We confirmed the PGC identification with periodic acid-Schiff (PAS) staining, anti-SSEA1, ß-catenin, ß-integrin, and Nanog immunofluorescence staining. Exogenously IDE1 treated-PGCs migrated toward the embryonic gonads after transplantation into the heart of the recipient embryo at HH stages 13-15. Our results suggested that the application of IDE1 small molecule into the culture of quail PGCs represented a step toward achieving efficient expansion of the avian PGCs.

Efficiency of early, single-dose probiotic administration methods on performance, small intestinal morphology, blood biochemistry, and immune response of Japanese quail.

The aim of the present study was to investigate the efficacy of early probiotics (single dose) administered in different ways, on quails' performance, small intestine morphology, blood biochemistry, and immune response. In total, 192 day-old chicks were used in one of the following experimental groups before being transferred to a raising room: 1) Control (no probiotic administered), 2) oral gavage, 3) spray, and 4) vent lip. Four replicates of 12 chicks per cage were considered for each treatment and birds were raised up to 35 d in the same conditions. Probiotic treated birds had higher d 1 to 35 feed intake than the control group (P < 0.05). In addition, oral-gavaged birds had a higher body weight gain as compared to the control (P < 0.05). The values of duodenum length and villus height of the oral group and ileum length and villus height of the vent lip group were greater than that of the control (P < 0.01). Regardless of the method of administration, probiotics resulted in deeper crypts and in a higher number of goblet cells in the duodenum and ileum as compared to the control (P < 0.01). The administration of probiotics resulted in increased plasma uric acid (P < 0.05), glucose, and total protein (P < 0.01). The concentration of hemoglobin was slightly higher in probiotic-supplemented groups. While a decreased concentration of triglyceride was observed in vent-lip probiotic-administered birds compared to control (P < 0.05), the concentration of cholesterol was not significantly affected by treatments (P > 0.01). None of the immune-related parameters were affected by the probiotic (P > 0.05). Single dose usage of probiotics exerts its beneficial effects on quails' body weight gain, feed intake and mortality in 1 to 35 d period, regardless of the route of administration. This work generally supports the efficacy of single-dose usage of probiotics and suggests the spray of probiotics as an early, single-dose administration method.