Chicken Intestine Research Articles

Technology Development to Suppress the Campylobacter Jejuni in Intestinal Microflora of The Broilers by Lactobacilli

This article presents a method for determining the microflora of the blind processes of the intestines of broiler chickens receiving heterologous lactobacilli with anti-C.jejuni activity as a feed additive in comparison with intact chickens. In the course of a scientific and production experiment on the Cobb 500 cross broilers, an experiment was carried out to determine the total number of bacteria in the intestinal contents by quantitative PCR. Sequencing was performed on a MiSeq genomic sequencer (Illumina, Inc., USA) using the MiSeq Reagent Kit v3 kit (Illumina, Inc., USA). Data were obtained on the total number of bacteria and their taxonomic diversity. As a result of the studies, it was shown that the microflora of broilers throughout the entire maintenance period corresponded to the norm. The use of feed additives based on lactobacilli in general had a positive effect on the state of microflora of the blind processes of the intestine..

Screening of Host Specific Lactic Acid Bacteria Active Against Escherichia coli From Massive Sample Pools With a Combination of in vitro and ex vivo Methods.

A novel three-step combination of in vitro and ex vivo screening was established to massively screen host derived lactic acid bacteria (LAB) from the broiler chicken intestine with inhibitory activity against Escherichia coli. In a first step, a massive sample pool consisting of 7102 broiler-derived colonies from intestinal contents were established and sub-cultured. Supernatants thereof were incubated with an E. coli model strain to screen suitable isolates with inhibitory activity. A total of 76 isolates of interest were subsequently further studied based on either pH dependent or -independent activity in the second step of the assay. Here, in-depth growth inhibition of the E. coli model strain and the potential of isolates for lactic acid production as inhibitory substance were indexed for all isolates. Resulting scatter plots of both parameters revealed five isolates with exceptional inhibitory activity that were further studied under ex vivo condition in the third step of the assay. These isolates were taxonomically classified as strains of the species Lactobacillus agilis, Lactobacillus salivarius, and Pediococcus acidilactici. Samples from the broiler chicken intestine were inoculated with the Lactobacillus isolates and the E. coli model strain. After 8 and 24 h incubation, respectively, growth of the E. coli model strain was monitored by cultivation of the E. coli strain in antibiotic supplemented medium. By their superior inhibitory activity against the E. coli model strain, one L. agilis and one L. salivarius strain were selected and characterized for further application as probiotics in broiler chicken. Additionally, their antibiotic resistance patterns and resilience under gastric stress of isolates were also characterized. The results of this study demonstrate that the novel isolation procedure was able to efficiently and rapidly isolate and identify bacterial strains from a massive sample pool with inhibitory potential against specific types of bacteria (here E. coli). The introduction of the final ex vivo selection step additionally confirmed the inhibitory activity of the strains under conditions simulating the intestinal tract of the host. Furthermore, this method revealed a general potential for the isolation of antagonistic strains that active against other pathogenic bacteria with specific biomarker.

PROX1 is a specific and dynamic marker of sacral neural crest cells in the chicken intestine.

The enteric nervous system (ENS) is a complex network constituted of neurons and glial cells that ensures the intrinsic innervation of the gastrointestinal tract. ENS cells originate from vagal and sacral neural crest cells that are initially located at the border of the neural tube. In birds, sacral neural crest cells (sNCCs) first give rise to an extramural ganglionated structure (the so-called Nerve of Remak [NoR]) and to the pelvic plexus. Later, sNCCs enter the colon mesenchyme to colonize and contribute to the intrinsic innervation of the caudal part of the gut. However, no specific sNCC marker has been described. Here, we report the expression pattern of prospero-related homeobox 1 (PROX1) in the developing chick colon. PROX1 is a homeobox domain transcription factor that plays a role in cell type specification in various tissues. Using in situ hybridization and immunofluorescence techniques, we showed that PROX1 is expressed in sNCCs localized in the NoR and in the pelvic plexus. Then, using real-time quantitative PCR we found that PROX1 displays a strong and highly dynamic expression pattern during NoR development. Moreover, we demonstrated using in vivo cell tracing, that sNCCs are the source of the PROX1-positive cells within the NoR. Our results indicate that PROX1 is the first marker that specifically identifies sNCCs. This might help to better identify the role of the different neural crest cell populations in distal gut innervation, and consequently to improve the diagnosis of diseases linked to incomplete ENS formation, such as Hirschsprung's disease.

Evaluation of effects of a novel probiotic feed supplement on the quality of broiler meat.

Background and Aim:A local microorganism-based probiotic has been developed as an alternative to using antibiotic growth promoter and its effect on broiler meat quality has been studied, when supplemented with poultry feed at different concentrations. This study aimed to understand the effect of local microorganism-based probiotic sourced from cattle rumen and chicken intestine and added as feed supplement at different concentrations on broiler meat quality.Materials and Methods:The local microorganism-based probiotic made from cattle rumen and chicken intestine contained Lactobacillus spp., Bifidobacterium spp., Streptococcus spp., and Bacillus spp. The experiments followed a completely randomized design. Treatments in the study were: P0, i.e., control (without probiotic), P1 (probiotic administered at 5 ml/kg feed), P2 (probiotic administered at 10 ml/kg feed), and P3 (probiotic administered at 15 ml/kg feed). Each treatment was repeated 5 times. Parameters examined in this study were pH, meat tenderness, fat content, and meat protein content.Results:Based on a total of 200 chickens, the percentage of meat protein content in treatments P1, P2, and P3 showed an increase of 19.34%, 19.42%, and 19.64%, respectively, when compared with P0 that showed a protein content increase of 19.14%. The fat content of meat for P1, P2, and P3 was 21.54%, 21.46%, and 21.30%, respectively, which was less than the value for P0 (21.69%). The treatments did not significantly affect pH or meat tenderness when compared with the control. The usage of this novel probiotic as a feed supplement resulted in an increase in meat protein content and a decrease in fat content.Conclusion:This study indicates that using the local microorganism-based probiotic sourced from cattle rumen and chicken intestine to supplement poultry feed did not have a significantly different effect (p>0.05) on meat pH; however, it had a significantly different (p<0.05) on protein and fat content of broiler meat.

Chicken nugget nutrition composition with an additional variation of breadfruit flour

The study aims to evaluate nuggets substituted with chicken intestine and breadfruit flour. The treatment consisted of 100% topica flour (P1), and substitution with breadfruit flour each of 15% (P2), 25% (P3), 35% (P4), 50% (P5), 65% (P6), 75% (P7), 85% (P8), 100% (P9). The study showed that the water content of P4 nugget to 100% (P9) was significantly lower than the control (P1). 100% ash content substitution nugget (P9) breadfruit flour was significantly higher than the control. The substitution of breadfruit flour does not affect protein and fat. It was concluded that the substitution of breadfruit flour with tapioca in the manufacture of the nugget was significantly different in water content and ash, but protein and fat content of the resulting nugget were relatively the same. The composition of 50% substitution nugget tapioca flour and breadfruit 50% (P5) contains 61.5% water, 1.65% ash, 17.55% protein, and 17.72% fat.

In vitro characterization and analysis of probiotic species in the chicken intestine by real-time polymerase chain reaction

Two experiments, 1 in vitro and 1 in vivo study, were conducted to analyze probiotic species characteristics and survival in the intestine of broiler birds. The in vitro study characterized the effect of bile salt supplementation and pH on the proliferation of Lactobacillus reuteri, Pediococcus acidilactici, Bifidobacterium animalis, and Enterococcus faecium. L. reuteri and P. acidilactici growth was maximal when the media was supplemented with 1.0% bile salt, whereas B. animalis and E. faecium growth was maximal when the media was supplemented with 0.5% bile salt. Altering the pH between 2.5 and 5.8 did not significantly (P > 0.05) alter the proliferation of L. reuteri and B. animalis. Decreasing the pH from 5.8 to 2.0 decreased P. acidilactici growth, whereas it increased the E. faecium proliferation. The in vivo study quantified the concentration of L. reuteri, P. acidilactici, B. animalis, E. faecium, and L. salivarius in different intestinal sections from birds supplemented with and without synbiotic containing the above 5 bacteria species. Birds were supplemented with and without synbiotic for 18 d, after which all birds were fed the same basal diet with no synbiotic. At 72h of feeding, the basal diet with no synbiotics, when the probiotic species in the feed is expected not to confound the recovery of probiotic species from the intestine, intestinal contents were collected. L. reuteri, P. acidilactici, E. faecium, and L. salivarius were below detectable amount in the control group. L. reuteri concentration expressed as copy numbers/g and as percentage of total bacteria was highest in the jejunum and ileum, respectively. E. faecium concentration was highest in the ileum. The copy number of P. acidilactici increased at the duodenum and plateaued after duodenum. L. salivarius concentration was highest in the jejunum. It can be concluded that real-time PCR can be applied to quantify the concentrations of probiotic species in the intestine and probiotic species differ in their ability to colonize different sections of the intestine.

Effect of different bait on the catchment of eels Anguilla marmorata in the Brayeun River, Aceh Besar district, Indonesia

The objective of the present study was to determine the best bait to catch the eel Anguilla marmorata in the Brayeun River, Aceh Besar District, Indonesia. Five types of baits were tested, namely; shrimps, earthworms, chicken intestine, terrestrial snails, and tilapia fish fingerling. The fishing was conducted for three months based on the Islamic Hijri Calendar (lunar cycle) on Rajab to Ramadhan (April to June 2017). The data were subjective to one-way analysis of variance (one-way ANOVA). A total of 39 eels A. marmorata were sampled during the study. The ANOVA test showed that the baits gave a significant effect on the catch volume of eel where the shrimp bait resulted in higher catching compared to other baits. Therefore, it is concluded that the shrimp was the best bait for eel A. marmorata in the Brayeun River, Aceh Besar District, Indonesia.

Molecular cloning, tissue distribution and the expression of cystine/glutamate exchanger (xCT, SLC7A11) in different tissues during development in broiler chickens.

The cystine/glutamate exchanger (xCT, SLC7A11) is a component of the system Xc amino-acid antiporter that is able to export glutamate and import cysteine into cells. The xCT amino acid exchanger has received a lot of attention, due to the fact that cysteine is an essential substrate for the synthesis of glutathione (GSH), an endogenous antioxidant in cells. The objective of this research was to clone the full-length cDNA of chicken xCT, and to investigate the gene expression of xCT in different tissues, including intestinal segments of broiler chickens during development. The full-length cDNA of chicken xCT (2,703 bp) was obtained from the jejunum by reverse transcription-PCR and sequenced. Homology tests showed that chicken xCT had 80.4%, 80.2%, and 71.2% homology at the nucleotide level with humans, cattle, and rats, respectively. Likewise, amino acid sequence analysis showed that chicken xCT protein is 86.4%, 79.3%, and 75.6% homologous with humans, cattle, and rats, respectively. Additionally, phylogenetic analysis indicated that chicken xCT genes share a closer genetic relationship with humans and cattle, than with rats. The chicken xCT protein has 12 transmembrane helixes, 6 extracellular loops, and 5 intracellular loops. The mRNA of xCT was detected in all tissues, including intestinal segments, in which the mRNA expression of xCT was significantly higher (P < 0.05) within the colon, compared to the jejunum and ileum. During development, a linear pattern of changes regarding the levels of the xCT mRNA was found, indicating that there was an abundance of xCT within the duodenum (P < 0.05). Furthermore, there were changes of the xCT mRNA abundance in the colon during development, which displayed linear and cubic patterns (P < 0.05). These results indicated that xCT is widely expressed both in intestinal segments, as well as other organs that are not associated with nutrient absorption. Further investigation is needed to characterize the functional relevance of xCT activity in oxidative stress and inflammation in the small intestine of broiler chickens.

Comparative genome analysis reveals metabolic traits associated with probiotics properties in Lactobacillus reuteri KUB-AC5

Lactobacillus reuteri KUB-AC5 is a heterofermentative lactic acid bacteria that was isolated from the chicken intestine. Due to L. reuteri KUB-AC5 has been often used as probiotics, there is still largely unknown on its genes and functions in relation to probiotic properties. This study therefore aimed to perform a genome sequencing of L. reuteri KUB-AC5 and further accomplished comparative analysis with overall identifying its unique/shared metabolic genes and functions across L. reuteri genomes. Accordingly, a genome sequence of the L. reuteri KUB-AC5 has a total length of 2,187,995 base pairs with a GC content of 39.29%. Based metabolic annotation, out of 2196 protein-encoding genes, 477 genes were mostly found to be involved in the metabolism, especially in carbohydrates, cofactors and vitamins. Of these, interestingly, cobalamin (vitamin B12) and folate (vitamin B9) were proposed to be in relation to probiotic properties of L. reuteri KUB-AC5. Considering gene clusters identification, putative unique exopolysaccharide (EPS) biosynthetic gene cluster was also identified in this KUB-AC5 strain. In context of bacteriocin-producing strain, the L.reuteri KUB-AC5 genome promisingly displays to have its genes involved in functional roles of immunity, regulator and transporter. Our study serves for guiding on how L. reuteri KUB-AC5 is used as probiotics for impact to host health. This study shows a broader impact for designing efficient probiotics for industrial foods and feeds.

Antagonistic trait of Lactobacillus reuteri S5 against Salmonella enteritidis and assessment of its potential probiotic characteristics

Salmonella enteritidis is an important foodborne pathogen that has caused multiple outbreaks of infection associated with poultry and egg consumption. Thus, the prevention and inhibition of Salmonella enteritidis infection are of great concern. Lactic acid bacteria have anti-pathogenic activity; however, their underlying mechanisms and modes of action have not yet been clarified. In this study, the antibacterial mechanism of Lactobacillus reuteri S5 (L. reuteri S5) against Salmonella enteritidis ATCC13076 (S. enteritidis ATCC13076) was studied by different methods. We found that L. reuteri S5 was able to form a stable biofilm formation, colonizing the entire intestinal tract of chickens. In addition, bacterial cultures and the cell-free supernatant (CFS) of L. reuteri S5 inhibited SE ATCC13076 growth, and this growth inhibition was also observed in the co-culture assay. This effect may be predominantly caused by antimicrobial metabolites produced by L. reuteri S5. Furthermore, treatment with the CFS of L. reuteri S5 resulted in a significant reduction in the expression of Salmonella virulence, motility and adhesion genes and a significant reduction in the motility ability and inhibitory effect on biofilm formation. In addition, the damage to the membrane structure and intracellular structure induced by the CFS of L. reuteri S5 could be observed on Transmission electron microscopy images and dodecyl sulfate, sodium salt (SDS)-Polyacrylamide gel electrophoresis confirmed the disruptive action of the CFS of L. reuteri S5 on the cytoplasmic membrane. Our findings demonstrate that L. reuteri S5, an intestinal Lactobacillus species associated with chicken health, is able to form biofilm and stably colonize chicken intestines. It also possesses anti-SE activity, preventing SE growth, inhibits the expression of SE genes involved in adhesion and invasion, virulence and cell membrane integrity, inhibits SE biofilm formation and motility, damages or destroys bacterial structures, and inhibits intracellular protein synthesis. L. reuteri S5 therefore has potential applications as a probiotic agent.

Microstructure of the small intestine in broiler chickens fed a diet with probiotic or synbiotic supplementation.

The aim of the study was to determine the effect of dietary supplementation of a probiotic and a synbiotic on the morphometric parameters of the small intestine of broiler chickens. The experiment was conducted on three hundred sixty, one-day-old female Ross 308 chicks, which were randomly selected from 20,000 birds and divided into three treatment groups (n=120) with ten replicates per treatment. The control group (C) was fed a commercial diet, the probiotic group (PRO) was fed the same diet with an added 1% of the probiotic Lavipan® (Lactococcus lactis, Carnobacterium divergens, Lactobacillus casei, Lactobacillus plantarum and Saccharomyces cerevisiae), and the synbiotic group (SYN) was fed the commercial diet with an added synbiotic: 0.8% of the prebiotic RFO (extracted from lupin seeds) and 1% Lavipan®. According to the manufacturer's data, apart from the typical probiotic action,microorganisms contained in the preparation release anti-bacterial substances (hydrogen peroxide and bacteriocins) and, therefore, are antagonistic towards pathogenic bacteria present in the gut of animals. Supplementation took place in the first seven days of rearing, and all birds had ad libitum access to water and feed during the 42days of the experiment. On the last day, all birds were slaughtered and samples from three segments of the small intestine were taken. Villi area, height, width and crypt depth ratios were read using Multiscan software. Synbiotic supplementation increased the BWG of broilers from first to tenth day of rearing, compared to the control group. The PRO group had improved villi morphometric parameters of the duodenum. In the jejunum and ileum, both bioactive substances improved villus width and villus surface area. Crypts were deeper in the small intestine of birds supplemented with bioactive substances, which allows greater renewal of the villi. As expected, the intestinal morphometric parameters of broiler chickens benefited from bioactive substance supplementation.

Effect of piperine and quercetin alone or in combination with marbofloxacin on CYP3A37 and MDR1 mRNA expression levels in broiler chickens

After oral route of administration, drug absorption is unpredictable and is governed by various factors such as multi drug resistance-1 (MDR1) an efflux transporter and drug metabolizing enzymes (like CYP3A4, CYP3A37, CYP2D6) at intestine and liver. Naturally available phyto chemicals like piperine and quercetin as well as some floroquinolones are known to inhibit MDR1 and CYP3A37 activity and increases bioavailability of co-administered drugs. This study was carried out to investigate the effect of piperine and quercetin alone or in combination with marbofloxacin on CYP3A37 and MDR1 mRNA expression levels in liver and intestine of broiler chicken. After oral administration of piperine and quercetin for 3 consecutive days followed by with or without oral administration of marbofloxacin for 5 days, CYP3A37 and MDR1 mRNA expression levels were determined using quantitative real-time PCR. Total of 36 broiler chickens in seven individual groups were treated with different regimen and the mRNA expression levels at duodenum and liver were analyzed with apt statistical tools. After piperine and quercetin combined treatment with marbofloxacin, CYP3A37 mRNA expression levels were significantly down regulated by 20.57 (p = .034) and 25.95 (p = .003) folds; and MDR1 mRNA expression levels were also significantly down regulated by 11.33 (p = .012) and 33.59 (p = .006) folds in liver and duodenum, respectively. Down regulation of CYP3A37 and MDR1 mRNA in liver and duodenum indicate the combined pretreatment of piperine and quercetin may be useful for improving the pharmacokinetics of orally administered drugs which are substrates for CYP3A37 and MDR1.

Antibiotic Resistance of Escherichia Coli Isolated from Broiler Chickens

Abstract The purpose of this study was to detect the antibiotic resistance of forty-one Escherichia coli isolates from the intestinal contents of slaughtered broiler chickens using the disk diffusion method according to Kirby-Bauer. Mueller-Hinton agar plates were inoculated with 0.1 ml overnight broth cultures of individual E. coli isolates and the disks with the following concentrations of antibiotics were applied onto them: ampicillin (10 μg), cefotaxime (30 μg), gentamicin (10 μg), streptomycin (10 μg), azithromycin (15 μg), tetracycline (30 μg), ciprofloxacin (30 μg) and levofloxacin (3 μg). After the incubation at 37 °C for 16—18 hours, the inhibition zones were measured and interpreted in accordance with the Clinical and Laboratory Standard Institute (CLSI) zone diameter breakpoints. Almost all E. coli isolates showed resistance to tetracycline (92.68 %), most of them were resistant to gentamicin (75.61 %) and levofloxacine (70.73 %). Phenotypic resistance to tetracycline was further confirmed with the help of the Polymerase Chain Reaction (PCR) procedure focused on the presence of specific tet(A) and tet(B) genes. These genes were detected in all 41 E. coli isolates. On the contrary, E. coli isolates were highly susceptible to both azithromycin and streptomycin. In conclusion, the study highlighted the role of commensal E. coli bacteria isolated from the intestines of broiler chickens as an important reservoir of tetracycline resistance genes.

Investigating the role of interleukin 10 on Eimeria intestinal pathogenesis in broiler chickens

Eimeria species are intestinal protozoan parasites that cause lack of production, malabsorption and mortality in floor raised chickens. Administering an oral antibody to interleukin 10 (aIL-10) reduces the symptoms of coccidiosis in broilers, indicating interleukin 10 (IL-10) is key to Eimeria pathology. IL-10 is an anti-inflammatory cytokine and acts as a stand down signal to reduce inflammation and host pathology during disease. Related protozoan parasites exploit IL-10 to reduce pathogen-damaging host inflammatory responses. We hypothesize that IL-10 is increased during Eimeria infection through an unknown host-pathogen interaction, and by feeding aIL-10 to neutralize excess IL-10 the bird is allowed to mount an effective immune response to Eimeria. To determine the effects of aIL-10 during the intestinal immune response, intestinal pathology and the relationship between IL-10, interferon gamma (IFNγ) and Eimeria infection were evaluated in this study.In both experiments, broilers were administered either a 10x dose of Advent® Eimeria vaccine or saline. Duodenum, jejunum and cecum samples were collected, processed, stained and examined under a microscope. Evaluation of intestinal histomorphology during aIL-10 administration showed minimal differences in birds fed aIL-10 during infection compared to animals fed a control antibody during Eimeria infection. To further evaluate aIL-10’s positive effect during infection, immunofluorescent histochemistry was performed on chicken intestines days 3-7 post Eimeria infection for IL-10 and IFNγ presence in intestinal mucosa in control and infected birds, in regions with and without visible Eimeria burden. IL-10 and IFNγ had significant changes between days 4.5-7 post-infection in birds fed aIL-10 compared to animals fed a control antibody. Overall we found that the duodenum had increased IL-10 presence and increased IFNγ presence, and the jejunum and cecum had decreased IL-10 presence and decreased IFNγ presence. These differences in spatial regulation of IL-10 and IFNγ may indicate Eimeria species induce slightly different cytokine responses.

Molecular characterization of surface antigen 10 of Eimeria tenella.

Chicken coccidiosis is caused by the apicomplexan parasite Eimeria spp. At present, drug resistance of Eimeria is common because of the indiscriminate use of anticoccidial drugs. The gene encoding surface antigen 10 of Eimeria tenella (EtSAG10) is differentially expressed between drug-resistant and drug-sensitive strains. RNA-seq analysis indicated that this gene was downregulated in strains resistant to maduramicin and diclazuril compared to susceptible strains. EtSAG10 DNA sequence alignment revealed that they contained one and ten mutations in MRR and DZR, compared with DS, respectively. A full-length EtSAG10 cDNA was successfully cloned and expressed, and the polyclonal antibody was prepared. The transcription and translation levels of EtSAG10 were analyzed by quantitative real-time PCR (qPCR) and Western blotting. The localization of EtSAG10 in Spz, Mrz, and parasites in the first asexual stage was determined by indirect immunofluorescence. The potential association of EtSAG10 with sporozoite invasion of host cells was assessed by invasion inhibition assays. The results showed that EtSAG10 had a predicted transmembrane domain at the C-terminal end and a predicted signal peptide at the N-terminal end. EtSAG10 was downregulated in drug-resistant strains, which is consistent with the RNA-seq results. The EtSAG10 protein was localized to the parasite surface and parasitophorous vacuole membrane. This protein was shown to play a role in the infection of chicken intestine by sporozoites.

Population pharmacokinetics for danofloxacin in the intestinal contents of healthy and infected chickens.

Avian pathogenic Escherichia coli could cause localized and systemic infection in the poultry, and danofloxacin is usually used to treat avian colibacillosis through oral administration. To promote prudent use of danofloxacin and reduce the emergence of drug-resistant E.coli strains, it is necessary to understand the population pharmacokinetics (PopPK) of danofloxacin in chicken intestines. In this study, reversed-phase high performance liquid chromatography (HPLC) with fluorescence detection was used to detect the concentrations of danofloxacin in the contents of duodenum, jejunum, and ileum of the healthy and infected chickens after single oral administration (5mg/kg body weight). Then, the PopPK of danofloxacin in intestines were analyzed using NONMEM software. As a result, a two-compartment PK model best described the time-concentration profile of duodenal, jejunal, and ileal contents. Interestingly, absorption rate (Ka ), distribution volume (V), and clearance (CL) for danofloxacin from duodenal, jejunal to ileal contents were sequentially decreased in the healthy chickens. However, the trend of Ka , V, and CL of danofloxacin was changed dramatically in the intestine of infected chickens. Ka and V of danofloxacin in the jejunum were higher than in the ileum and duodenum. Compared with healthy chickens, Ka and V of danofloxacin in the duodenum decreased significantly, while increased in jejunum, respectively. It has been noted that Ka decreased and V increased in the ileum of infected chickens. Besides, CL in the duodenum, jejunum, and ileum of infected chickens was, respectively, lower than those of healthy chickens. Interestingly, the relative bioavailability (F) of danofloxacin in the ileum was relatively higher in both healthy and infected chickens. In addition, F in the duodenal, jejunal, and ileal contents of infected chickens was respectively higher than healthy chickens. In summary, the PopPK for danofloxacin in infected chicken intestines was quite different from healthy chickens. The absorption, distribution, and clearance of danofloxacin in healthy chickens decreased from duodenum to jejunum and to ileum. Moreover, the pharmacokinetic characteristics in the intestine of infected chickens changed significantly, and the pharmacokinetic characteristics in the ileum can be used as a representative of all intestinal segments.

The design of a cell-selective fowlicidin-1-derived peptide with both antimicrobial and anti-inflammatory activities

Fowlicidin-1 (Fowl-1), a cathelicidin expressed in chicken intestine, is known to have both antimicrobial and anti-inflammatory properties. However, its pharmaceutical development has been ultimately compromised by its high host cytotoxicity. In this study, a series of N- and C-terminal-truncated 19-meric Fowl-1 peptides were synthesized. Among these truncated peptides, Fowl-1 (8–26) exhibited broad-spectrum antimicrobial activity without human erythrocyte cytotoxicity while reducing anti-inflammatory activity. Further, Fowl-1 (8–26)-WRK was designed via Thr5→Trp, Ile7→Arg, and Asn11→Lys substitutions in Fowl-1 (8–26) to exhibit more amphipathicity. The results revealed that it exhibited both antimicrobial and anti-inflammatory properties. This study also demonstrated that the inhibitory activity of Fowl-1 (8–26)-WRK against LPS-induced inflammation was mainly due to the binding of LPS to the peptide. Interestingly, compared with human cathelicidin LL-37 and melittin, Fowl-1 (8–26)-WRK showed more potent activity against drug-resistant bacteria. It was also resistant to physiological salts and human serum and acted synergistically in combination with conventional antibiotics, such as chloramphenicol, ciprofloxacin, and oxacillin, suggesting that combined with conventional antibiotics, it is a promising adjuvant. Furthermore, membrane depolarization, SYTOX Green uptake, and flow cytometry revealed that it kills bacteria by damaging their membrane integrity. Therefore, this study suggests that Fowl-1 (8–26)-WRK has considerable potential for future development as an antimicrobial and anti-inflammatory agent for treating antibiotic-resistant infections.

In vivo and in vitro protective effect of arginine against intestinal inflammatory response induced by Clostridium perfringens in broiler chickens

BackgroundNecrotic enteritis is a widespread disease in poultry caused by Clostridium perfringens. We previously reported that dietary arginine supplementation protected the intestinal mucosa of broiler chickens with necrotic enteritis, but the related protective mechanisms remain unclear. The in vivo trial was designed as a 2 × 2 factorial arrangement to evaluated the effects of arginine supplementation on inflammatory responses, arginine transporters, arginine catabolism and JAK-STAT signalling pathway in broiler chickens challenged with C. perfringens or without C. perfringens. Furthermore, we validated the in vivo results using intestinal epithelial cells of chicken embryos.ResultsC. perfringens infection markedly increased gut gross pathological and histopathological lesion scores, promoted liver C. perfringens invasion, reduced serum arginine levels, and elevated jejunal mucosal lysozyme activities (P < 0.05), but these effects were significantly reversed by arginine supplementation in vivo (P < 0.05). The challenge significantly increased serum procalcitonin levels, jejunal mucosal iNOS activities and jejunal IL-6, TGF-β3, cationic amino acid transporter (CAT)-1, and CAT-3 mRNA expression (P < 0.05), whereas arginine supplementation significantly reduced jejunal IFN-γ, IL-1β, IL-6, IL-10, TGF-β3, and CAT-3 mRNA expression (P < 0.05). Arginine supplementation significantly attenuated the C. perfringens challenge-induced increases in jejunal iNOS, arginase 2, arginine decarboxylase, arginine:glycine amidinotransferase, JAK1, JAK3, STAT1, and STAT6 mRNA expression (P < 0.05). The in vitro experiment showed that C. perfringens challenge markedly increased cellular cytotoxicity and the mRNA expression of IL-1β, IL-8, IL-10, CAT-1 and CAT-3 (P < 0.05), which were significantly reversed by 50 μmol/L and/or 400 μmol/L arginine pre-treatment (P < 0.05).ConclusionsArginine prevented C. perfringens challenge-induced circulated arginine deficiency, normalized intestinal arginine transport and catabolism, down-regulated JAK-STAT signalling pathway and attenuated the inflammatory response, which exerted protective effects on the intestine of broiler chickens.

The influence of prophylactic biopreparations on preservation and microbiocenosis of chickens

The cultivation of organic birds is an important stage in ensuring the population, in particular children, a source of quality and easily digestible protein. The use of antibiotics with a prophylactic purpose in organic livestock is prohibited at the legislative level. Therefore, this new direction in agriculture requires professional scientific support and effective natural prophylaxis as an alternative to antibiotics. The research was conducted in a certified organic poultry farm. The purpose of the production trial of the probiotic and the new postbiotic drug on chickens was to compare their effectiveness with respect to the productivity and consistency of the poultry. The results were compared with the data of the control group of chickens and experimental groups among themselves. In addition, there was a link between the performance of chickens with the functional state of intestinal microbiocenosis and the corrective effect of these drugs. The expressed prophylactic effect of the tested drugs on the best of 20–22% preservation of chickens, as well as on higher body weight gain was established. The death of the control group chickens was due to dysbiosis of digestion with the domination of E. coli in the intestine. The study of intestinal microbiocenosis showed significantly higher concentrations of lactobacillus in the chickens of both experimental groups compared with the control group. Due to the competitive substitution of lactobacilli, the amount of E. coli in the intestines of chickens in experimental groups was reduced, where the test drugs were used. Excessive E. coli in the intestines of the control group chickens could provoke inflammation in the intestine and the development of clinical manifestations of colibacillosis. As a consequence – recorded a high percentage of deaths of chickens in this group. Thus, correction of microendocology of poultry intestines may be carried out by prophylactic preparations based on living representatives of symbiotic microflora (probiotics) or compositions of their useful metabolites (postbiotics). Such a correction occurs naturally, without violating the traditional mechanisms of interaction within the microbiological pool.

Scientific Coformer Screening, Preparation and Evaluation of Fenofibrate Tartaric Acid Cocrystal

Objective: This present study aims to screen pharmaceutical cocrystal of Fenofibrate and coformers. Further the preparation and evaluation of fenofibrate-coformer cocrystal and In-Vitro drug release and Ex-Vivo Permeation study was done. Material and Methods: The coformers for Fenofibrate were screened using molecular docking. The cocrystals produced were characterized using Differential Scanning Calorimetry (DSC), X-ray diffraction (XRPD) study and Infrared spectroscopy. Results: Cocrystal of Fenofibrate with tartaric acid was successfully prepared. The cocrystals displayed enhanced dissolution rate by 2.36 fold, similarly the ex-vivo drug uptake through everted chicken intestine model was improved by 4.38 fold. The formation of cocrystals of fenofibrate with tartaric acid was evaluated by DSC, IR and XRPD. Conclusion: The fenofibrate - tartaric acid cocrystal exhibited increased % drug release and permeation compared to fenofibrate. This study confirms that selection of proper coformer is very vital step in preparation of stable, superior cocrystal. Based upon above study and results it revealed that cocrystallization offers a valuable way to improve the physicochemical properties of the API. Keywords: Pharmaceutical Cocrystal, Fenofibrate , Coformer, Molecular docking.