Effect of sage (Salvia officinalis L.) extract in antioxidant status and intestinal morphology of pulmonary hypertensive chickens.

OBJECTIVES: The effects of dietary sage on the growth performance, antioxidant status, intestinal mucosa morphology, and pulmonary hypertensive response were investigated in broiler chickens with pulmonary hypertension. METHODS: Chicks (Ross 308) were reared under cold stress for 35 days and treated with 0.05% vitamin C (positive control) and 0 (control), 0.1 or 0.2% sage extracts, then performance, oxidant and antioxidant status, and intestinal morphology were evaluated. RESULTS: The index of pulmonary hypertension (RV:TV) was decreased, and weight gain (days 22-35) was increased in all treatments (except for sage 0.1%) compared with control (P < 0.05). Lipid peroxidation was decreased, whereas the activity of antioxidant enzymes (GPX, CAT, and SOD) was increased in the sage 0.2% group compared with control (P < 0.05). In the lung, SOD, CAT, and GPX transcripts were decreased in the sage 0.2% group compared with control (P < 0.05). In the right ventricle of the heart, SOD and CAT transcripts were increased in the sage 0.2% group compared with other groups of chickens, whereas GPX transcript was decreased (P < 0.05). The jejunal villus length in the chickens fed sage was significantly lower than in control (P < 0.05). The ileal villus width, villus surface area, and lamina proporia thickness in the chickens fed sage (0.2%) were increased compared with control (P < 0.05). CONCLUSIONS: Dietary supplementation of sage (0.2%) could modulate pulmonary hypertensive response, improve antioxidant status (enzymatic activity), intestinal morphometry, and absorptive surface in the broiler chickens.

Effect of Shirazi thyme on oxidant status and absorptive surface area of the intestine in cold-induced pulmonary hypertensive broiler chickens.

The effects of Shirazi thyme as a medicinal plant on oxidant status (lipid peroxidation, protein oxidation, total antioxidant capacity, and catalase activity) and absorptive surface area were measured in three segments of the small intestine in cold-induced pulmonary hypertensive chickens. Birds were reared at four groups (thyme 0, 0.25, 0.50, and 1.00 % of diet) for 42 days. To induce pulmonary hypertension, the temperature was gradually decreased. The body weight was increased in thyme-0.25% birds compared to control ones, while it was decreased in thyme-1% of birds. The feed consumption was only increased in thyme-1.00% birds. The feed conversion ratio was lower in thyme-0.25% birds and higher in thyme-1.00% birds than control ones. The duodenal and jejunal villus surface area was lower in thyme-1.00% birds than control ones, while it was greater in the thyme-0.50% birds. The ileal villus surface area and duodenal lamina propria thickness were also greater in thyme-0.50% birds. Lipid peroxidation was only decreased in the duodenum and ileum of thyme-0.50% birds compared to control ones, whereas it was increased in the duodenum and jejunum of thyme-1.00% birds. Catalase activity was only elevated in the duodenum and jejunum of thyme-1.00% fed chickens. Total antioxidant capacity was increased in the duodenum, jejunum, and ileum of thyme-0.50% birds. It is concluded that the Shirazi thyme has beneficial effects on growth performance, intestinal absorptive surface area / secretory system, and pulmonary hypertension response at low doses (0.25 and 0.50% of diet), whereas high dose (1.00% of diet) of this plant may be toxic.

Identification of reliable reference genes for quantitative real-time PCR in ovary and uterus of laying hens under heat stress.

The main stage in real-time quantitative PCR is a quantification of gene transcriptomes, in which suitable use of reliable reference genes is critical to normalize accurately. To determine the most stable reference genes in laying hens under heat stress, from a panel of nine typical candidate reference genes, the mRNA transcript of ACTB, HMBS, HPRT1, RPL13, RPL32, 18SrRNA, TBP, TFRC, and YWHAZ was evaluated in the ovary and uterus of both control and heat-stress groups of laying hens. Forty 23-week-old White Leghorn laying hens were housed in two rooms. The control (n = 20) and heat-stress (n = 20) groups were maintained at 21-23 °C and 36-38 °C for 8 weeks respectively. Analysis of this set of genes was done with BestKeeper, geNorm, and NormFinder software programs to find the most stable ones. Candidate reference genes ranked in the uterus of heat-stress and control groups of hens included YWHAZ, HPRT1, HMBS, RPL13, TFRC, ACTB, TBP, RPL32, and 18SrRNA; those in the ovary were YWHAZ, HPRT1, TFRC, HMBS, RPL13, TBP, RPL32, ACTB, and 18SrRNA. The overall results indicated that the most stable genes are YWHAZ, HPRT1, HMBS, RPL13, TFRC, TBP, ACTB, RPL32, and 18SrRNA respectively. In addition, the combination of YWHAZ, HPRT1, and HMBS is suggested as the most stable reference group of genes for more accurate quantitative data normalization in the ovarian and uterine tissues of laying hens under control and heat stress conditions. Lay summary Heat stress influences the expression of many genes in the reproductive tissues of birds. Accurate evaluation of these changes via real-time quantitative PCR depends on the determination of reliable reference genes. In this study, nine candidate housekeeping genes were evaluated, and the most stable were YWHAZ, HPRT1, HMBS, RPL13, TFRC, TBP, ACTB, RPL32, and 18SrRNA.

Effect of heat stress on the gene expression of ion transporters/channels in the uterus of laying hens during eggshell formation.

Heat stress is a problem in laying hens as it decreases egg quality by decreasing eggshell mineralization. Heat stress alters gene expression, hence our aim was to investigate effects of heat stress on gene expression of ion transport elements involving in uterine mineralization (TRPV6, CALB1, ITPR3, SCNN1G, SLC4A4, KCNJ15, SLC4A9, and CLCN2) by real time quantitative PCR. Forty 23-week-old White Leghorn laying hens were housed in two rooms. The control group (n = 20) was maintained at 21-23 °C, and the heat stress group (n = 20) was exposed to 36-38 °C for 8 weeks. All parameters of egg quality including egg weight, surface area, volume, and eggshell weight, thickness, ash weight, and calcium content were decreased in the heat stress group compared to the control group (by 26.9%, 32.7%, 44.1%, 38.4%, 31.7%, 39.4%, and 11.1%, respectively). Total plasma calcium was decreased by 13.4%. Levels of ITPR3, SLC4A4, and SLC4A9 transcripts in the uterine lining were decreased in the heat stress group compared to the control group (by 61.4%, 66.1%, and 66.1%, respectively). CALB1 transcript level was increased (by 34.2 fold) in the heat stress group of hens compared to controls. TRPV6, SCNN1G, KCNJ15, and CLCN2 transcript levels did not significantly differ between control and heat stress groups of laying hens. It is concluded that the down-expression of ITPR3, SLC4A4, and SLC4A9 genes may impair transportation of Cl-, HCO3-, and Na+ in eggshell mineralization during heat stress. Increased CALB1 gene expression may increase resistance of uterine cells to detrimental effects of heat stress.

Oxidant and enzymatic antioxidant status (gene expression and activity) in the brain of chickens with cold-induced pulmonary hypertension.

To evaluate oxidant and antioxidant status of the brain (hindbrain, midbrain, and forebrain) in chickens with cold-induced pulmonary hypertension, the measurements of lipid peroxidation, protein oxidation, antioxidant capacity, enzymatic activity, and gene expression (for catalase, glutathione peroxidase, and superoxide dismutases) were done. There were high lipid peroxidation/protein oxidation and low antioxidant capacity in the hindbrain of cold-induced pulmonary hypertensive chickens compared to control (P < 0.05). In the hypertensive chickens, superoxide dismutase activity was decreased (forebrain, midbrain, and hindbrain), while catalase activity was increased (forebrain and midbrain) (P < 0.05). Glutathione peroxidase activity did not change. Relative gene expression of catalase and superoxide dismutases (1 and 2) was downregulated, while glutathione peroxidase was upregulated in the brain of the cold-induced pulmonary hypertensive chickens. Probably, these situations in the oxidant and antioxidant status of the brain especially hindbrain may change its function at cardiovascular center and sympathetic nervous system to exacerbate pulmonary hypertension.

Effects of benzalkonium chloride on planktonic growth and biofilm formation by animal bacterial pathogens.

BACKGROUND: Resistance toward quaternary ammonium compounds (QACs) is widespread among a diverse range of microorganisms and is facilitated by several mechanisms such as biofilm formation. OBJECTIVES: In this study, the effects of benzalkonium chloride on planktonic growth and biofilm formation by some field isolates of animal bacterial pathogens were investigated. MATERIALS AND METHODS: Forty clinical isolates of Escherichia coli, Salmonella serotypes, Staphylococcus aureus and Streptococcus agalactiae (10 isolates of each) were examined for effects of benzalkonium chloride on biofilm formation and planktonic growth using microtiter plates. For all the examined strains in the presence of benzalkonium chloride, biofilm development and planktonic growth were affected at the same concentrations of disinfectant. RESULTS: The means of strains growth increase after the minimal inhibitory concentration (MIC) were significant in all the bacteria (except for E. coli in 1/32 and S. agalactiae in of 1/8 MIC). Biofilm formation increased with decrease of antiseptics concentration; a significant increase was found in all the samples. The most turbidity related to S. aureus and the least to Salmonella. CONCLUSIONS: Bacterial resistance against quaternary ammonium compounds is increasing which can increase the bacterial biofilm formation.

Chlorhexidine digluconate effects on planktonic growth and biofilm formation in some field isolates of animal bacterial pathogens.

BACKGROUND: To study chlorhexidine digluconate disinfectant effects on planktonic growth and biofilm formation in some bacterial field isolates from animals. OBJECTIVES: The current study investigated chlorhexidine digluconate effects on planktonic growth and biofilm formation in some field isolates of veterinary bacterial pathogens. MATERIALS AND METHODS: Forty clinical isolates of Escherichia coli, Salmonella serotypes, Staphylococcus. aureus and Streptococcus agalactiae (10 isolates for each) were examined for chlorhexidine digluconate effects on biofilm formation and planktonic growth using microtiter plates. In all of the examined strains in the presence of chlorhexidine digluconate, biofilm development and planktonic growth were affected at the same concentrations of the disinfectant. RESULTS: Chlorhexidine digluconate inhibited the planktonic growth of different bacterial species at sub-MICs. But they were able to induce biofilm development of the E. coli, Salmonella spp., S. aureus and Str. agalactiae strains. CONCLUSIONS: Bacterial resistance against chlorhexidine is increasing. Sub-MIC doses of chlorhexidine digluconate can stimulate the formation of biofilm strains.

Comparison of the anticoccidial effect of granulated extract of Artemisia sieberi with monensin in experimental coccidiosis in broiler chickens.

Coccidiosis is the most important parasitic disease of the poultry production industry. Due to increasing resistance to conventional anticoccidial agents, it is necessary to find new anticoccidial compounds. Herbal compounds such as those from Artemisia species are promising weapons in this regard since preliminary studies have shown its anticoccidial effects. To compare the anticoccidial effect of a granulated extract of Artemisia sieberi (GEAS) versus monensin in experimental broiler coccidiosis, 120 one-day old Ross 308 broiler chickens were divided in four groups, each with three replicates (n=10). Group 1 was separated as an uninfected negative control and received no treatment. At 21days of age, groups 2, 3 and 4 were inoculated with a mixed suspension of 2×10(5) oocysts of Eimeria tenella, Eimeria maxima, Eimeria acervulina and Eimeria necatrix. Group 2 was maintained as an infected positive control and received no treatment while groups 3 and 4 received GEAS (5mg/kg feed), and monensin (110mg/kg feed) from the first day until 42days of age as a feed additive, respectively. Five days after inoculation, the number of oocycts per gram (OPG) of feces for 7 successive days was measured. Also, mean body weight (MBW), weight gain (WG), feed intake (FI) and feed conversion ratio (FCR) were determined weekly in all groups and replicates. The results showed that GEAS and monensin improved performance attributes (FI, MBW, WG, FCR) and significantly (P<0.05) decreased OPG in inoculated broiler chickens.