Chicken Kidney Research Articles

Mitophagy Induced by Mitochondrial Function Damage in Chicken Kidney Exposed to Cr(VI).

Cr(VI) is a heavy metal environmental pollutant and carcinogen. Excessive Cr(VI) exposure injures kidneys. This study aimed to investigate mitophagy induced by mitochondrial function damage in chicken kidney exposed to Cr(VI). To explore the mechanism involved, we randomly divided 40 one-day-old Hy-line Brown cockerels into four groups, with each group exposed to different concentrations of Cr(VI), i.e., 0, 10, 30 and 50mgkg-1, which were orally administered daily for 45days. Excessive Cr(VI) increased tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and chemokine (C-X-C motif) ligand 1(CXCL1) expression and decreased Ca2+-adenosine triphosphatase (Ca2+-ATPase), Mg2+-ATPase and Na+/k+-ATPase activities in chicken kidney. Furthermore, Cr(VI) significantly increased reactive oxygen species (ROS) production and induced mitochondrial membrane potential (MMP) collapse and typical autophagosome formation. With the increase of Cr(VI) concentration, the Parkin translocation, value of LC3-II increased and decreased the content of p62/SQSTM1 and the translocase of outer mitochondrial membrane 20 (TOMM20). In summary, our findings explicated that mitochondrial function damage and mitophagy-related indicators were related to Cr(VI) concentration in chicken kidney.

Ecotoxicity of trace elements to chicken GALLUS gallus domesticus exposed to a gradient of polymetallic-polluted sites.

Mining activity may cause heavy metal accumulation, which threatens human and animal health by their long-term persistence in the environment. This study aims to assess the impact of polymetallic pollution on chicken (Gallus domesticus) from old lead mining sites in northeast of Tunisia: Jebel Ressas (JR). Samples of soil and chickens were collected from five sites being ranked along a gradient of heavy metal contamination. Heavy metal loads were evaluated in soil samples and in chicken liver and kidney. Biochemical evaluation of oxidative stress parameters termed as Catalase (CAT), Glutathione-S-Transferase (GST), and Malondialdehydes (MDA) accumulation was monitored. Metallothionein protein level was assessed as a specific response to heavy metals. DNA alteration was achieved using MNi frequency in the investigated tissues. Finally, the evaluation of gene expression levels of CAT, GST, mt1, mt4, P53, bcl2, caspase3 and DNA-ligase was performed. Our data showed the highest loads of Cd, Cu, Zn and Pb in tissues of animals from site 3, being more pronounced in kidney. Biochemical data suggested a significant increase in antioxidant enzymes activities in all sites respect to control except in site 3 were CAT and GST were inhibited. DNA alteration was observed in all tissues being very pronounced in animals from site 3. Overall, transcriptomic data showed that genes involved in apoptosis were up-regulated in animals exposed to the most contaminated soils. Our data suggest that chicken and selected biomarkers offer a suitable model for biomonitoring assessment of heavy metals transfer through the food web in mining sites. Finally, the obtained results of heavy metals accumulation and related alterations should be carefully considered in view of the controversial relationship between distribution and toxicology of contaminants in exposed organisms.

Analysis of highly polar pesticides and their main metabolites in animal origin matrices by hydrophilic interaction liquid chromatography and mass spectrometry

In this work, a method for the analysis of 14 highly polar pesticides in animal origin matrices by hydrophilic interaction liquid chromatography and mass spectrometry has been described. Three methanol-based extraction procedures with slight modifications, depending on the matrix, have been used. The method has been validated according to the performance criteria from the SANTE guideline for pesticide residues analysis in food and feed. Most of the compounds showed a limit of quantitation at the lowest spike levels tested (0.01–0.02 mg kg−1) in the matrices evaluated. A SPE clean-up step with mixed mode cation exchange sorbent has been used with some matrices, like chicken meat, eggs, liver and kidney, in order to improve the limits of quantification. Glufosinate, glyphosate and HEPA were the compounds which presented more difficulties during the study, due to the lack of sensitivity of the qualifier ions and/or because of the interference from the matrices.

Disorders of the metabolic status and morphofunctional state of liver and kidneys of chicken

In this study we analyzed biochemical parameters of blood, tibia, histological analysis of liver and kidney tissues of 46‐days old pullets of egg‐laying chickens. The research aim is study of biochemical and morphological changes in replacement pullets to predict risks of growth weak replacement pullets. The research shows that in the serum total proteins were higher then normal range on 28, 8%, albumins on 32,0%. High level of this parameters leads to produce too much uric acid and accumulate it in the body and its higher on 21,3%. Energy change in serum is shown with satisfying range of glucose and reduced rate of total lipids. Sufficient calcium in serum 13,1% with lack of phosphorus. Ca/P ratio exceeds 1.2 times optimal value. Low alkaline phosphatase activity is typical for inadequate bone matrix formation. AST is higher on 25,1% AST and ALT parameters are both high on 25,1% and 15,4% respectively and it means liver function abnormal. There is very low vitamin B2 (42,4% from normal value) wwith sufficiently high carotene supply with vitamins A, E, C in the liver. Liver micronutrients: Zinc – 2,9 times more; Manganese – 3,1 times more with less concentration of Cobalt on 2,6 times and Selenium – 8,3 times. There are calcium 1,6 times less and phosphorus – 1,4 in tibia. Thus in the liver of chickens has focal or diffuse inflammation with blood flow disturbances and of necrosis were recorded. Macroscopic examination found that kidney are oversized, have flabby consistency with light brown color and surface look like almond nut. Microscopic examination found that Bowman's capsule of the cortex kidney is thickened, the space between the capsule and glomeruli is swollen, and the borders of the tubule epithelial cells are poorly distinguishable. The cells are swollen, enlarged and cone‐shaped into the gap make it narrow. The degree of swelling of the epithelial cells, and, consequently, the narrowing of the lumens were uneven. In some tubules, the gaps are still quite distinguishable, in others they look like gaps. There are tubules in which the gaps are almost invisible. Nuclei are not clearly expressed in all cells. The established violations in the interstitial metabolism and structural changes in the parenchymal organs (liver, kidneys) during the restoration of youth is a predisposing load that ensures the body's resistance to adverse environmental influences and loss of productive metabolism during egg production.Micromorphology of the chickens' liver. Impaired blood circulation. Hematoxylin and eosin.X 640Figure 1The architectonics of the glomerular zone of the kidney in chickens 46 days old. Hematoxylin and eosin.X 640Figure 2

Selenium Yeast Alleviates Ochratoxin A-Induced Apoptosis and Oxidative Stress via Modulation of the PI3K/AKT and Nrf2/Keap1 Signaling Pathways in the Kidneys of Chickens.

The purpose of this study was to investigate the protective effect and mechanism of yeast selenium (Se-Y) on ochratoxin- (OTA-) induced nephrotoxicity of chickens. A total of 80 one-day-old healthy chickens were randomly divided into 4 equal groups: control, OTA (50 μg/kg OTA), Se-Y (0.4 mg/kg Se-Y), and OTA+Se-Y (50 μg/kg OTA+0.4 mg/kg Se-Y). In the OTA chickens, differences in body weight, kidney coefficient, biochemical histological analysis, antioxidant capability, and the expression levels of the PI3K/AKT and Nrf2/Keap1 signaling pathway-related genes were observed. The levels of total superoxide dismutase (T-SOD), antioxidant capacity (T-AOC), catalase (CAT), and glutathione (T-GSH) significantly decreased, but the malondialdehyde (MDA) level of the kidneys significantly increased in the OTA treatment group. More importantly, treatment with Se-Y improved the antioxidant enzyme activities within the kidneys of chickens exposed to OTA. In addition, administration of OTA resulted in apoptosis and was associated with decreased expression of AKT, PI3K, and Bcl-2, which in turn enhanced expression of Caspase3, Bax, and P53. However, Se-Y improved the antioxidant defense system through activation of the Nrf2/Keap1 signaling pathway. Gene expression of Nrf2 and its target genes (HO-1, GSH-px, GLRX2, MnSOD, and CAT) was downregulated following OTA exposure. Conversely, Se-Y treatment resulted in a significant upregulation of the same genes. Besides, significant downregulations of protein expression of HO-1, CAT, MnSOD, Nrf2, and Bcl-2 and a significant upregulation of Caspase3 and Bax levels were observed after contaminated with OTA. Notably, OTA-induced apoptosis and oxidative damage in the kidney of chickens were reverted back to normal level in the OTA+Se-Y group. Taken together, the data suggest that Se-Y alleviates OTA-induced nephrotoxicity in chickens, possibly through the activation of the PI3K/AKT and Nrf2/Keap1 signaling pathways.

Recombinant infectious bronchitis coronavirus H120 with the spike protein S1 gene of the nephropathogenic IBYZ strain remains attenuated but induces protective immunity

Infectious bronchitis (IB) is a highly infectious viral disease responsible for major economic losses in the poultry industry. A reverse genetic vaccine is a safe, rapid, and effective method of achieving IB prevention and control. In this study, we constructed the recombinant strain, rH120-S1/YZ, using a reverse genetic system, based on the backbone of the H120 vaccine strain, with the S1 gene replaced with that of the QX-like nephropathogenic strain, ck/CH/IBYZ/2011, isolated in China. The results of dwarf chicken embryos, growth kinetics, and viral titration in the embryos demonstrated that the biological characteristics of the recombinant virus remained unchanged. Like the rH120-infected group and in contrast to the rIBYZ-infected group, no mortality, clinical signs, or lesions were observed in the lungs or kidneys of young chickens inoculated with rH120-S1/YZ. The viral loads in various tissues, cloacal, and oral swabs was lower in most types of samples, indicating that the rH120-S1/YZ strain was highly safe in chicks. Compared to rH120 vaccination group, when the efficacy of this strain was evaluated against the QX-like IBV strain, better protection, with 100% survival rate and no disease symptom or gross lesion was observed in the chickens vaccinated with rH120-S1/YZ. Increased levels of IBV-specific antibodies were detected in the serum of the rH120-S1/YZ-vaccinated animals 14 days post-vaccination. Collectively, our results suggest that the recombinant strain, rH120-S1/YZ, may represent a promising vaccine candidate against QX-like IBVs.

Gga-miR-30d regulates infectious bronchitis virus infection by targeting USP47 in HD11 cells

Avian infectious bronchitis virus (IBV) is a coronavirus which infects chickens and causes severe economic losses to the poultry industry worldwide. MicroRNAs (miRNAs) are important intracellular regulators and play a pivotal role in viral infections. In previous studies, we have revealed that IBV infection caused a significant down-regulation of gga-miR-30d expression in chicken kidneys. In present study, we investigated the role of gga-miR-30d in the process of IBV infection of HD11 cell line in vitro. By transfecting the mimics and inhibitor of gga-miR-30d, it was found that overexpressed gga-miR-30d inhibited IBV replication. Contrarily, low-expressed gga-miR-30d promoted IBV replication. In addition, dual-luciferase reporter assays revealed that ubiquitin-specific protease 47 (USP47), a deubiquitinase-encoding gene, was a target for gga-miR-30d. This is the first study demonstrating that miRNAs regulate IBV replication by regulating the deubiquitinating enzyme (DUBs).

Comparative histological changes in the structure of the spleen and kidneys of experimental chickens exposed to the action of “Guidamis” as an adjuvant for vaccination against infectious bronchitis

The article provides a histological description of the internal organs – the spleen and kidneys of chickens exposed to the action of “Guidamis” as an adjuvant. A lysate was tested as part of a sample of the vaccine PB-07 against infectious bronchitis developed by researchers of the Department of Virology and Tumor Diseases of Birds of the Russian Agricultural Academy (Lomonosov). The experiments were carried out on Lohmann Brown chickens in the conditions of the Laishev branch of “Ak Bars” Poultry Company, the Republic of Tatarstan. The prolonged action of the vaccine against infectious bronchitis and the use of “Guidamis” as an adjuvant was manifested as an increase in the proliferation and differentiation of lymphoid tissues in the numerous lymph nodes. The use of the vaccine and the adjuvant had a weak reactogenic effect. In the spleen of control chickens, hyperplasia of lymphoid tissue cells is weakly expressed. In the periarterial zone of nodules of the thymus-dependent lymphocytes, the cell density is low. In the kidneys, for fourteen days after the vaccination against infectious bronchitis using Guidamis, the moderate granular dystrophy of the epithelium, mainly of individual distal tubules, and local moderate manifestations of edema of the capsule of the renal corpuscles were observed. In the kidneys, signs of pronounced edema of the capsule cavity, significant disturbances in the patency of the tubular network caused by swelling and partial desquamation of the epithelium, as well as the presence of epithelium and protein mass in the lumen of the tubules, were observed.

Pathological effect of infectious bronchitis disease virus on broiler chicken trachea and kidney tissues.

Aim:This study aimed to investigate the pathological effects of the infectious bronchitis virus (IBV) on chicken trachea and kidney tissues and also desired to diagnose the virus genome using a molecular tool.Materials and Methods:Twenty trachea and kidney samples collected from one broiler farm contain 10,000 chickens at Tikrit city. The chickens showed signs of gasping and mortality (20%) at early ages (20 days old), the presence of IBV investigated using conventional reverse transcriptase-polymerase chain reaction technique with routine histopathological study to tracheal and renal tissue.Results:Postmortem lesion showed severe respiratory inflammation with abscesses at tracheal bifurcation lead to airway blog. Molecular results showed two genotypes of IBV, one of them not included in primer designer research. The histological study showed different stages of inflammation, degeneration, and necrosis to the renal and tracheal tissues.Conclusion:The respiratory and renal pathological effect of the virus responsible for the symptoms appeared on the affected chicks that caused mortality, with a high probability of presence of a new viral genotype added to the untranslated region.

The Preparation of Chicken Kidney Cell Cultures for Virus Propagation.

Chicken kidney (CK) cells have been widely utilized in virus research studies for many years. The optimized technique of primary CK cell culture production involving both mechanical and enzymatic disaggregation is described. This updated method proved to consistently give high cell yields and resultant cultures are readily used for virus assays.

Adjuvant Effects of Platycodin D on Immune Responses to Infectious Bronchitis Vaccine in Chickens.

Adjuvants are common vaccine components. Novel adjuvants may improve the protective immunity conferred by vaccines against poultry diseases. Here, a less-hemolytic saponin, platycodin D (PD), isolated from the root of Platycodon grandiflorum was investigated as a potential alternative adjuvant. PD was tested as an adjuvant in the infectious bronchitis (IB) vaccine, because the existing IB vaccine has often failed to induce effective immune responses. The adjuvant activity of PD in conjunction with IB vaccine was evaluated in this study. Compared to control treatment, PD treatment significantly increased the proliferation of chicken peripheral blood mononuclear cells, concentration of interferon-γ in culture supernatants, and anti-IB antibody titer. In chickens pre-challenged with the Mass 41 infectious bronchitis virus (IBV), PD administration resulted in fewer and less severe clinical signs, lower mortality rate, and higher protection compared to control treatment. Histopathological examination showed that the lungs and kidneys of PD-treated chickens displayed fewer pathological lesions than those of control chickens. Our results also demonstrated that this new vaccine adjuvant improved chicken humoral and cellular immune responses without any side effects. Hence, our findings suggest that PD might serve as an effective adjuvant in IBV vaccines.

Effects of the Soil-Derived Microorganism BX-1 on Chicken Newcastle Disease

In recent years, effective microorganisms (EMs) have been administered to humans and domestic animals, and their usefulness has been recognized for promoting health and enhancing immunity. For example, the preventative effects against flu are enhanced by ingestion of Lactobacillus by humans, and symptom relief of atopic dermatitis has been reported, with EMs actually used in commercial products. In addition, EM preparations are being used in livestock to prevent infections (e.g. Salmonella and Escherichia coli infection). In poultry, avian influenza and Newcastle disease are terrible and fatal infectious diseases that cause significant economic damage. Furthermore, countries designated as contaminated with these pathogens can experience major trade problems. Given the above, how to protect livestock from infections safely and at low cost without using disinfectants, antibiotics and vaccines is a major issue. In the present study, we examined whether or not Newcastle disease could be suppressed by feeding chickens BX-1 as an EM feed. A field strain of Newcastle virus was cloned from cloaca swabs of large numbers of dying chickens in a poultry farm in Indonesia by polymerase chain reaction (PCR) and hemaggregation assays. Chicken kidney cells and embryonated eggs were highly sensitive to this virus, and high titers of virus were able to be collected. The experimental viral inoculated to chickens showed a high mortality rate, with high pathogenicity in birds. Conventional chickens were also raised on a diet supplemented with BX-1 and directly infected with the Newcastle virus. The mortality was decreased in these infected birds. Even the low dose of BX-1 had an inhibitory effect on the lethality of the infection. These results suggest that BX-1 intake through an EM diet is effective in controlling Newcastle disease.

Pathogenicity of a QX-like avian infectious bronchitis virus isolated in China

Avian infectious bronchitis is a serious and highly contagious disease caused by infectious bronchitis virus (IBV). We isolated a highly virulent IBV strain (CK/CH/JS/TAHY) from kidneys of diseased chickens. Phylogenetic analysis based on the S1 gene revealed that CK/CH/JS/TAHY clustered with the QX-like type. The S1 gene has 1,620 nucleotides and encoded a polypeptide of 540 amino acids with typical coronavirus cleavage recognition sites of HRRR. About 1-day-old specific pathogen-free White Leghorn chickens inoculated with CK/CH/JS/TAHY at 105.5 EID50 exhibited clinical signs including coughing, sneezing, nasal discharge, and tracheal vocalization accompanied by depression with 84% mortality and 100% morbidity. The kidneys of dead birds were swollen and pale and exhibited severe urate deposition. Histopathological examination revealed kidney hemorrhages, multifocal necrosis of the renal tubules and trachea with cilia loss, sloughing of epithelial cells, and edema of the lamina propria. IBV-specific antibodies appeared at 10 D post-infection. Chickens vaccinated with a CK/CH/JS/TAHY oil-emulsion vaccine showed 26.7% morbidity and 3% mortality indicating a protective effect. In conclusion, the IBV strain is a virulent avian IBV and that exhibited severe pathogenicity in chickens and is a vaccine candidate to prevent infection by Chinese QX-like nephropathogenic IBV strains.

Dietary Betaine Improves Intestinal Barrier Function and Ameliorates the Impact of Heat Stress in Multiple Vital Organs as Measured by Evans Blue Dye in Broiler Chickens.

Simple SummaryHeat stress alters the normal physiological status, compromising the function of organs such as the small intestine. However, evidence exists of a wider distribution of organ dysfunction, stemming from factors such as a reduction in blood flow due to redistribution to the skin for increased radiant heat loss to the environment. Simultaneously, assessing organ dysfunction at multiple locations presents technical difficulties, and hence studies are lacking. Therefore, the aim of this experiment was to determine the pattern of Evans Blue Dye distribution as a cost-effective indicator of organ dysfunction in HS chickens supplemented with betaine. The results showed that Evans Blue Dye concentration increased in the kidney and muscle during heat stress, while such concentration was reduced with betaine. Therefore, betaine could improve the broiler’s tolerance to heat stress, and Evans Blue Dye may be a useful tool for investigating the effects of heat stress on broiler organ dysfunction.In a 2 × 2 factorial design, 60 male Ross-308 broilers were fed either a control or 1 g/kg betaine diet and housed under thermoneutral (TN) or heat stress (HS) conditions. Broilers were acclimated to diets for 1 week under TN (25 °C), then either kept at TN or HS, where the temperature increased 8 h/day at 33 °C and 16 h/day at 25 °C for up to 10 days. Respiration rate (RR) was measured at four time points, and on each of 1, 2, 3, 7 and 10 days of HS, 12 broilers were injected with 0.5 mg/kg of Evans Blue Dye (EBD) solution to quantify regional changes in tissue damage. Betaine was quantified in tissues, and ileal damage was assessed via morphometry and transepithelial resistance (TER). Heat stress elevated RR (p < 0.001) and resulted in reduced villous height (p = 0.009) and TER (p < 0.001), while dietary betaine lowered RR during HS (p < 0.001), increased betaine distribution into tissues, and improved ileal villous height (p < 0.001) and TER (p = 0.006). Heat stress increased EBD in the muscle and kidney of chickens fed the control diet but not in those receiving betaine. Overall, these data indicate that supplemented betaine is distributed to vital organs and the gastrointestinal tract, where it is associated with improved tolerance of HS. Furthermore, EBD markers help reveal the effects of HS on organs dysfunction.

Detection of Norfloxacin in Muscle, Liver and Kidney of Broiler Chicken

The present study was undertaken to determine the residues of norfloxacin in muscle, liver and kidney of broiler chicken. On the basis of surveillance study, commonly used antibiotic norfloxacin was selected for quantifying the concentration in broiler chicken meat samples. The HPLC system was equipped with photodiode array and C18 hypersil column. The mobile phase consisting of 1 ml Ortho phosphoric acid (85 per cent v/v), 100 ml water HPLC grade with acitonitrile HPLC grade, 87:12 (v/v) then pH was adjusted to 2.0 with triethylamine HPLC Grade. The results indicated that out of 180 samples analysed, 3 liver samples of target stations, viz. T3, T6 and T10 were found positive for norfloxacin residues with concentrations of 2.00, 2.60 and 2.31 µg/kg, respectively. Samples collected from other target stations did not show residual concentration of norfloxacin in muscle, liver and kidney samples.

Denatonium Benzoate-Induces Oxidative Stress in the Heart and Kidney of Chinese Fast Yellow Chickens by Regulating Apoptosis, Autophagy, Antioxidative Activities and Bitter Taste Receptor Gene Expressions.

Simple SummaryDenatonium benzoate is a strong bitter taste receptor agonist, extensively used for its activation of different cell pathways. Taste signals have been associated to food recognition and avoidance, and bitter taste provokes an aversive reaction and is assumed to protect chickens from consuming poisons and harmful toxic substances. The results of the study revealed that dietary supplementation with medium and high doses of denatonium benzoate damaged the epithelial cells of the heart and kidneys by inducing apoptosis and autophagy and reduced the growth of chickens, respectively. However, mRNA expressions of bitter taste receptors, downstream signaling effector genes, apoptosis-, autophagy- and antioxidant-related genes were higher on day 7, while these expressions were subsequently decreased on day-28 in the heart and kidney of Chinese Fast Yellow chickens in a dose-response manner.The sense of taste which tells us which prospective foods are nutritious, poisonous and harmful is essential for the life of the organisms. Denatonium benzoate (DB) is a bitter taste agonist known for its activation of bitter taste receptors in different cells. The aim of the current study was to investigate the mRNA expressions of bitter taste, downstream signaling effectors, apoptosis-, autophagy- and antioxidant-related genes and effector signaling pathways in the heart/kidney of chickens after DB dietary exposure. We randomly assigned 240, 1-day-old Chinese Fast Yellow chicks into four groups with five replicates of 12 chicks and studied them for 28 consecutive days. The dietary treatments consisted of basal diet and feed containing DB (5, 20 and 100 mg/kg). The results revealed that dietary DB impaired (p < 0.05) the growth performance of the chickens. Haemotoxylin and eosin staining and TUNEL assays confirmed that medium and high doses of DB damaged the epithelial cells of heart/kidney and induced apoptosis and autophagy. Remarkably, the results of RT-PCR and qRT-PCR indicated that different doses of DB gradually increased (p < 0.05) mRNA expressions of bitter taste, signaling effectors, apoptosis-, autophagy- and antioxidant- related genes on day 7 in a dose-response manner, while, these expressions were decreased (p < 0.05) subsequently by day-28 but exceptional higher (P < 0.05) expressions were observed in the high-dose DB groups of chickens. In conclusion, DB exerts adverse effects on the heart/kidney of chickens in a dose-response manner via damaging the epithelium of the heart/kidney by inducing apoptosis, autophagy associated with bitter taste and effector gene expressions. Correlation analyses for apoptosis/autophagy showed agonistic relationships. Our data provide a novel perspective for understanding the interaction of bitter taste, apoptosis, autophagy and antioxidative genes with bitter taste strong activators in the heart/kidney of chicken. These insights might help the feed industries and pave the way toward innovative directions in chicken husbandry.

Milk thistle protective effect on liver and kidney of broiler chickens

Milk thistle, Silybum marianum is a useful medicinal plant, whose extract has antioxidant properties. The protective effect of the milk thistle medicinal plant was studied on the liver and kidney of Ross broiler chickens. Milk thistle at three levels (0%, 0.3%, 3%) as well as cotrimoxazole at two levels (0 g/kg, 2 g/kg) were used in five replications. Blood metabolites were measured at age 42 d. Data were analyzed using a general linear model. The effect of milk thistle levels was significant(p < 0.05) on blood cholesterol. Cotrimoxazole had a highly significant (p < 0.01) effect on blood cholesterol. The effect of milk thistle levels was significant (p < 0.05) on the levels of uric acid and blood creatinine. The effect of milk thistle levels was highly significant (p < 0.01) on blood bilirubin. The difference in the mean levels of milk thistle and cotrimoxazole was significant (p < 0.05) for prothrombin time. The use of different levels of milk thistle reduced liver and kidney problems and modified the undesirable effects of cotrimoxazole. A level of 3% milk thistle had the most protective effect on the liver and kidney of broilers.

Comparative Analysis of Gene Expression in Virulent and Attenuated Strains of Infectious Bronchitis Virus at Subcodon Resolution.

Like all coronaviruses, avian infectious bronchitis virus (IBV) possesses a long, single-stranded, positive-sense RNA genome (∼27 kb) and has a complex replication strategy that includes the production of a nested set of subgenomic mRNAs (sgmRNAs). Here, we used whole-transcriptome sequencing (RNASeq) and ribosome profiling (RiboSeq) to delineate gene expression in the IBV M41-CK and Beau-R strains at subcodon resolution. RNASeq facilitated a comparative analysis of viral RNA synthesis and revealed two novel transcription junction sites in the attenuated Beau-R strain, one of which would generate a sgmRNA encoding a ribosomally occupied open reading frame (dORF) located downstream of the nucleocapsid coding region. RiboSeq permitted quantification of the translational efficiency of virus gene expression and identified, for the first time, sites of ribosomal pausing on the genome. Quantification of reads flanking the programmed ribosomal frameshifting (PRF) signal at the genomic RNA ORF1a/ORF1b junction revealed that PRF in IBV is highly efficient (33 to 40%). Triplet phasing of RiboSeq data allowed precise determination of reading frames and revealed the translation of two ORFs (ORF4b and ORF4c on sgmRNA IR), which are widely conserved across IBV isolates. Analysis of differential gene expression in infected primary chick kidney cells indicated that the host cell response to IBV occurs primarily at the level of transcription, with global upregulation of immune-related mRNA transcripts following infection and comparatively modest changes in the translation efficiencies of host genes. Cellular genes and gene networks differentially expressed during virus infection were also identified, giving insights into the host cell response to IBV infection.IMPORTANCE IBV is a major avian pathogen and presents a substantial economic burden to the poultry industry. Improved vaccination strategies are urgently needed to curb the global spread of this virus, and the development of suitable vaccine candidates will be aided by an improved understanding of IBV molecular biology. Our high-resolution data have enabled a precise study of transcription and translation in cells infected with both pathogenic and attenuated forms of IBV and expand our understanding of gammacoronaviral gene expression. We demonstrate that gene expression shows considerable intraspecies variation, with single nucleotide polymorphisms being associated with altered production of sgmRNA transcripts, and our RiboSeq data sets enabled us to uncover novel ribosomally occupied ORFs in both strains. The numerous cellular genes and gene networks found to be differentially expressed during virus infection provide insights into the host cell response to IBV infection.

Responsiveness Expressions of Bitter Taste Receptors Against Denatonium Benzoate and Genistein in the Heart, Spleen, Lung, Kidney, and Bursa Fabricius of Chinese Fast Yellow Chicken.

Simple SummaryIn chickens, bitter taste is the most significant biological taste disrupter; it is believed to protect chickens against consuming poisonous/toxic materials and considered a warning signal prior to ingestion. The bitter taste receptors’ extraoral expression information is deficient in chicken, and denatonium benzoate is extensively used as a bitter taste receptor agonist in different cells. Our results found that qRT-PCR showed a high level of dose-dependent expressions of ggTas2Rs in the starter and grower stages in the heart, spleen, lungs, and kidneys, while the dose-dependent expressions were lower in the bursa Fabricius. The growth performance of the selected organs significantly (and unexpectedly) improved upon the administration of denatonium benzoate 5 mg/kg and genistein 25 mg/kg treatments, while the gains in organ weights were impaired in the groups given denatonium benzoate 20 mg/kg and 100 mg/kg, respectively.The present study was conducted to investigate the responsiveness expressions of ggTas2Rs against denatonium benzoate (DB) and genistein (GEN) in several organs of the Chinese Fast Yellow Chicken. A total of 300 one-day-old chicks that weighed an average of 32 g were randomly allocated into five groups with five replicates for 56 consecutive days. The dietary treatments consisted of basal diet, denatonium benzoate (5 mg/kg, 20 mg/kg, and 100 mg/kg), and genistein 25 mg/kg. The results of qRT-PCR indicated significantly (p < 0.05) high-level expressions in the heart, spleen, and lungs in the starter and grower stages except for in bursa Fabricius. The responsiveness expressions of ggTas2Rs against DB 100 mg/kg and GEN 25 mg/kg were highly dose-dependent in the heart, spleen, lungs, and kidneys in the starter and grower stages, but dose-independent in the bursa Fabricius in the finisher stage. The ggTas2Rs were highly expressed in lungs and the spleen, but lower in the bursa Fabricius among the organs. However, the organ growth performance significantly (p < 0.05) increased in the groups administered DB 5 mg/kg and GEN 25 mg/kg; meanwhile, the DB 20 mg/kg and DB 100 mg/kg treatments significantly reduced the growth of all the organs, respectively. These findings indicate that responsiveness expressions are dose-dependent, and bitterness sensitivity consequently decreases in aged chickens. Therefore, these findings may improve the production of new feedstuffs for chickens according to their growing stages.

Large-scale potential RNA editing profiling in different adult chicken tissues.

RNA editing is a post-transcription maturation process that diversifies genomically encoded information and can lead to transcriptome diversity. Thanks to next-generation sequencing technologies, a large number of editing sites have been identified in different species. Although this mechanism is well described in mammals, only a few studies have been performed in chicken. Here, candidate or potential RNA editing sites were identified in eight different tissues of chicken (brain, spleen, colon, lung, kidney, heart, testes and liver). We identified 68 A-to-G editing sites in 46 genes. Only two of these were previously reported in chicken. We found no C-to-T sites, attesting to the lack of this type of editing mechanism in chicken. Similar to mammals, the editing sites were enriched in non-coding regions, rarely resulted in a change in amino acids, showed a critical role in the nervous system and had a low guanosine level upstream of the editing site and some enrichment downstream from the site. Moreover, in contrast to mammals, editing sites were weakly enriched in interspersed repeats and the number and editing ratio of non-synonymous sites were higher than for those of synonymous sites. Interestingly, we found several tissue-specific edited genes, including GABRA3, SORL1 and HTR1D in brain and RYR2 and FHOD3 in heart, that were associated with functional processes relevant to the corresponding tissue. This finding highlights the importance of RNA editing in several chicken tissues, especially the brain, and establishes a foundation for further exploration of this process.