14th Day Of Incubation Research Articles

THE FATTY ACIDS CONTENT IN THE LIVER OF JAPANESE QUAILS AFTER THE CHEMICAL TREATMENT OF HATCHING EGGS

The five groups of Pharaoh quail (100 animals per group) were formed to fulfill the tasks. Quails of the control group were fed by the standard mixed fodder for quail (DSTU 4687:2006). The bird of the research groups (I–IV) received the same feed, but with the addition of 20 g/t of vitamin E. We selected the hatching eggs after 4 weeks of feeding research fodders. Eggs were weighed and laid for incubation using the standard mode after pre-incubation storage of the eggs of the quails obtained in the peak of egg production for 5 days. On the 14th day of incubation, the eggs of the quails were divided into 5 groups. Treatment of eggs I, II and III of the test group was carried out on the 14th day of incubation, respectively, with solutions of 1% sodium hypochlorite 2% perchloric acid 0,5% hydrogen peroxide. Egg of control and IV experimental group were not subject to chemical treatment. The material for research was hatching eggs of quail, liver tissue of 14 daily embryos and 1-day quail. The fatty acid composition of the lipids of tissues and egg yolk was determined in this biological material. Analysis of methyl esters of LC was carried out on a Gas chromatograph Trace GC Ultra (USA) with a flame ionization detector. Identification of fatty acids was carried out using a standard Supelco 37 Component FAME Mix. Quantitative assessment of the spectrum of fatty acids of yolk lipids was carried out by the method of internal normalization, determining their percentage content. Ontogenetic differences in the fatty acid composition of tissues are established. Processing of the eggshell on the fourteenth day of incubation with H2O2 solution is accompanied by an increase in the proportion of stearic acid in the liver of the 1-day quail and a decrease in the proportion of Neukosatrienoic and pre-fatty acids. Treatment of the shell with sodium hypochlorite and HCl is accompanied by an increase in the proportion of stearic acid by 0.96–1.00 % and аrachinic acid, with a decrease in the proportion of gondoinic and eicosatrienic fatty acids. At the same time, when treating with HCl and sodium hypochlorite, the ratio of the sum of saturated to unsaturated fatty acids decreases by 3.2–7.9% (p<0.05). So, the established changes in the fatty acid composition of the liver one-day quail indicate a significant effect of the chemical treatment of the egg shell on the exchange of fatty acids in the embryonic period.

Selected prebiotics and synbiotics administered in ovo can modify innate immunity in chicken broilers

BackgroundA previous study showed that prebiotics and synbiotics administered in ovo into the egg air cell on the 12th day of incubation enhance the growth and development of chickens. However, the influence of this procedure on the development and efficiency of the innate immune system of broiler chickens is unclear. Therefore, the aim of this study was to evaluate whether the early (on the 12th day of embryo development) in ovo administration of selected prebiotics (inulin − Pre1 and Bi2tos − Pre2) and synbiotics (inulin + Lactococcus lactis subsp. lactis IBB SL1 − Syn1 and Bi2tos + L. lactis subsp. cremoris IBB SC1 − Syn2) influences the innate immune system.ResultsChickens (broiler, Ross 308) that were treated with Pre1 exhibited a decreased H/L ratio on D7, but an increased H/L ratio was observed on D21 and D35. In the remaining experimental groups, an increase in the H/L ratio was observed on D21 and D35. The oxidative potential of leukocytes measured using the NBT test increased on D21 in Pre2 and Syn1 groups. The rate of the phagocytic ability of leukocytes increased in Pre1 and Syn1 groups on D21. The phagocytic index decreased in Pre1 and Syn2 groups on D21 and D35. Concurrently, the count of WBC in circulating blood decreased on D21 in Pre1, Pre2, and Syn1 groups. The hematocrit value was increased in Syn1 chickens on D21, in Pre1 chickens on D35, and in Syn2 chickens on both time points.ConclusionsEarly in ovo treatment of chicken embryos with prebiotics and synbiotics may temporarily modulate not only the production/maturation of leukocytes but also their reactivity.

EFFECT OF ESSENTIAL OILS OF LAMIACEAE PLANTS ON THE PENICILLIUM COMMUNE

The aim of this research was to determine the inhibitory effect of vapor phase of eight essential oils (EOs) on the growth of seven strains of Penicillium commune isolated from moldy milk products. Another objective was to determine the minimum inhibitory doses (in vitro and probit analyses) of EOs, which at concentration 625 μL.L-1 of air completely inhibited the growth of all strains. The antifungal activity was evaluated by the micro-atmosphere method. Thyme, red thyme, peppermint, mint, and savory completely inhibited the growth of strains during cultivation at 25 °C and 5 °C. Basil, rosemary, and sage EOs have different effects on the growth of P. commune strains. EOs that completely inhibit the growth of all strains were used to determine their minimum inhibitory doses (MIDs). The best results 62.5 μL.L-1 of air and 125 μL.L-1 of air 7th day at 25 °C of incubation showed red thyme EO. Similar results were also found in thyme EO, but the MID inhibiting the growth of the one strain was 250 μL.L-1 of air. MIDs of savory and peppermint were from 125 to 500 μL.L-1 of air. Mint EO had the highest MID (from 250 to 625 μL.L-1 of air). On the 14th day of incubation we found the same MIDs, respectively higher. It was found that EOs have different effects on individual strains of P. commune. According to probit analyses, the most effective tested EOs were red thyme and thyme, less effective peppermint and savory, and the least effective was mint EO.

Effects of In-Ovo Injection of Manganese on Some Organ Weights and Lengths in Quail Hatching Eggs

This study investigated the effects of in-ovo injection of manganese on some organ weights and length in quail hatching ( Coturnix coturnix japonica ) eggs. In total, 400 quail hatching eggs were randomly divided into four treatment groups of 100 eggs per treatment with four replicates of 25 eggs each. On the 14th day of incubation, eggs from group 1 were not injected (control(C)), group 2 was injected with Mn-bioplex at 20 µg per egg (Mn20), group 3 was injected with Mn-bioplex at 40 µg per egg (Mn40), and group 4 was injected with Mn-bioplex at 80 µg per egg (Mn80). There were no significant differences among treatments for chick weight, yolk sac weight, liver weight, and heart weight. The highest chick length was obtained from the C treatment. No significant differences were found in chick length among Mn treatment groups. The beak length of C treatment were lower compares with that of the Mn80 treatment group (P<0.05). But no significant differences were observed in beak length among Mn treatments groups. The leg length in C group was significantly lower than Mn40 and Mn80 treatments groups, except Mn20 treatment group (P<0.01). As a result, it can be said that injection of Mn into the hatching quail eggs causes the chick and leg lengths to decrease, and beak length to increase without affecting the weight parameters examined.

Embryonic development of parakeratinized epithelium of the tongue in the domestic duck (Anas platyrhynchos f. domestica): LM, SEM, and TEM observations

The parakeratinized epithelium is a common and widespread type of keratinized epithelium in the oral cavity in adult birds. In contrast to orthokeratinized epithelium, which mostly covers mechanical papillae and the lingual nail, parakeratinized epithelium covers almost the entire dorsal surface of the tongue in birds. The characteristic feature of parakeratinized epithelium is the presence of nuclei in the keratinized layer. The present study aimed to investigate for the first time the micro- and ultrastructural changes of parakeratinized epithelium during embryonic development and to assess the readiness of the epithelium to serve protective functions during food transport to the esophagus. Three developmental stages were distinguished: embryonic, transformation, and pre-hatching stages. The embryonic stage lasts from the 9th to the 14th day of incubation and the epithelium is composed of undifferentiated epithelial cells. The transformation stage lasts from the 15th to the 22nd day of incubation and the epithelium undergoes transformation into stratified epithelium consisting of basal, intermediate, and superficial layers. The characteristic feature of this stage is formation of the periderm with osmophilic granules. The pre-hatching stage starts on the 23rd day, and the epithelium with a fully developed keratinized layer resembles that of the epithelium in adult animals. No periderm was observed on the epithelial surface. It was confirmed that at the time of hatching the parakeratinized epithelium is fully differentiated and ready to fulfill its function during food transport. The presence of periderm is a common feature characteristic for para- and orthokeratinized epithelium in the oral cavity of birds. However, the formation of the keratinized/cornified layer is different for these two types of keratinized epithelia.

Effect of cultural conditions on antimicrobial activity of marine-derived fungus Penicillium chrysogenum

Marine fungi are a potential source of structurally diversified bioactive secondary metabolites that are not found in terrestrial sources. In our continuous investigation to search new antimicrobial agents from marine-derived fungi, we isolated fungal strain 045-357-2 from a soft coral sample collected from Ca Na bay, Ninh Thuan, Vietnam. The strain showed high antibacterial activity and was selected for further study. According to Internal Transcribed Spacer (ITS) rDNA molecular methods, the fungus was identified as Penicillium chrysogenum 045-357-2 and had a 100% homology (600/600 bp) with a GenBank sequence from a reference P. chrysogenum strain (NCBI accession no. EF200090). In the present investigation, the fungus was studied on effect of different culture incubation period, pH and salinity for antimicrobial activity against pathogenic microbes including Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Streptococcus faecalis, Listeria monocytogenes, and Candida albicans. The ethyl acetate extracts of culture medium of the fungal isolate were determined antimicrobial activity by disc diffusion assay. The results showed that the optimum antimicrobial activity of the fungal strain P. chrysogenum 045-357-2 was obtained in medium containing 20 g rice, 20 mg yeast extract, 10 mg KH2PO4 , and 40 mL natural seawater at 30oC on the 14th day of incubation. The strain also exhibited highest antimicrobial activity in the medium at initial pH 6.0 – 8.0 and salinity of 35 g/L. The findings indicate that cultural conditions significantly influenced on antimicrobial activity of the studied fungus. The optimization is potential use for further study on the mass cultivation and isolation of bioactive compounds for this fungus.

Effects of urease and nitrification inhibitors on the soil mineral nitrogen dynamics and nitrous oxide (N2O) emissions on calcareous soil.

Urease inhibitors and nitrification inhibitors can reduce nitrogen (N) loss in agriculture soil. However, the effect of inhibitors on soil N2O emissions under the drip irrigation system remains unclear. A pot and a field experiment with two inhibitors were conducted to explore how inhibitors regulate soil nitrogen transformation and N2O emissions. In the pot experiment, three treatments included control, urea, and urea + N-(n-butyl)thiophosphoric triamide (NBPT, urease inhibitor). In the field experiment, three treatments included control, urea, and urea + NBPT + 2-chloro-6-(trichloromethyl)pyridine (nitrapyrin, nitrification inhibitor). The urease inhibition rate in the treatment of urea + NBPT was 27.5% at the 14th day of incubation (pot experiment), and NH4+-N was significantly decreased by 37-64% compared with urea alone treatment. In the field experiment, the nitrification inhibition rate in the treatment of urea + NBPT + nitrapyrin was 47.7 and 63.9% on the 3rd day after fertilization at the wheat heading and filling stages, respectively. Compared to urea treatment, NO3--N concentration in the double-inhibitor-added treatment was significantly decreased by 32 and 20% on the 5th day after fertilization at the heading and filling stages, respectively; N2O fluxes were also decreased by 30.9 and 33.3% at the two stages of wheat, respectively. In total, adding an inhibitor reduced N loss by 7.39 and 7.44% at the 14th and 35th day in the pot experiment and by 10.53 and 6.65% at the two growing stages of wheat in the field experiment, respectively. Path and correlation analysis showed that N2O emissions were significantly correlated with soil NO3- in both pot and field experiments.

Histomorphogenesis of pancreas in ostrich embryo (Struthio camelus)

The purpose of this study was evolution of pancreas in ostrich. The study was done on 24 incubated eggs of ostrich embryo on 10, 13, 16, 22, 26, 30 and 36 days of prehatching and 1 day of post hatching life. The histological stains such as haematoxylin eosin (H&E), gomori and Toluidine blue were used for this study. On 13th day of incubation the primary pancreas was developed near the liver and duodenum and it was composed of one part. The pancreas was consisted of undifferentiated epithelial cells, connective tissue, non -organized ducts and blood vessels. On 16th day the pancreas was composed of dorsal and ventral lobe with the similar structure to 13th day. On the 22th day, the zymogen granules were evident in developing exocrine part. By aging the level of connective tissue and ducts were decreased and the acinar cells were increased. On 1th day after hatching the hepatocytes vacuolated severely and pancreatic islets were appeared.

Short-term effects of tidal flooding on soil nitrogen mineralization in a Chinese tidal salt marsh

Tidal flooding is an important control of nitrogen biogeochemistry in wetland ecosystems of Yellow River Delta, China. Variations in hydrology could change soil redox dynamics and conditions for microorganisms living. A tidal simulation experiment was designed to extract tidal flooding effect on nitrogen mineralization of salt marsh soil. Inorganic nitrogen and relevant enzyme were measured during the 20-day incubation period. Considering the variation of both inorganic N and enzymes, nitrogen mineralization process in tidal salt marsh could be divided into 2 phases of short term response and longtime adaption by around 12th incubation day as the inflection point. Soil ammonium nitrogen (NH4+-N) and volatilized ammonia (NH3) occupied the mineralization process since nitrate nitrogen (NO3−-N) was not detected over whole incubation period. NH4+-N varied fluctuant and increased significantly after 12 day's incubation. Released NH3 reached to peak value of 14.24 mg m−2 d−1 at the inflection point and declined thereafter. Inorganic nitrogen released according to net nitrogen mineralization rate (RM) under the tidal flooding condition without plant uptake except first 2 days. However, during the transitional period of 6–12 days, RM decreased notably to almost 0 and increased again after inflection point with the value of 0.182 mg kg−1 d−1. It might be due to the change of microbial composition and function when soil shifted from oxic to anoxic, which were reflected by arylamidase, urease and fluorescein diacetate. Fluorescein diacetate hydrolysis and arylamidase had the similar variation of U style with decreasing activities before 12 days' incubation. All the enzymes measured in this experiment increased after inflection point. Whereas, urease activity kept constant from 2 to 12 days. Alternant oxidation reduction condition would increase N loss through denitrification and ammonia volatilization during the transitional period, while more inorganic nitrogen would be available in reductive environment of long-term tidal flooding. Therefore, hydrological process regulation has great influence on nitrogen cycling and further influence on wetland productivity.

MICROBIAL EXTRACTION OF CHITIN AND CHITOSAN FROM PLEUROTUS SPP, ITS CHARACTERIZATION AND ANTIMICROBIAL ACTIVITY

Objective: The aim of the present work was to extract the chitin and chitosan from two species of Pleurotus, its characterization and its antimicrobial activity against bacteria and fungi.Methods: The fungi were grown in MGYP medium for 15 d. The chitin and chitosan were extracted using the alkaline method and various biochemical methods and compared with commercial chitin and chitosan. The FTIR and SEM were also performed. Antimicrobial activity was performed using well diffusion method.Results: The maximum yield of mycelia and the chitin and chitosan were obtained on 12th day of incubation. P. folrida gave maximum yield (201.3 mg/l of chitin and 65.61 mg/l of chitosan). Fourier Transform Infrared spectroscopy comparing the results obtained with results for commercial chitin and chitosan. Our results showed that chitin and chitosan obtained from Pleurotus spp. had a significant similitude with commercial. SEM image also carried out to study the surface morphology. The fungal chitin and chitosan had more antimicrobial activity against the gram+ve, gram–ve and the various fungus.Conclusion: In the present study the chitin and chitosan were extracted from the Pleurotus spp. (P. florida and P. eous). The yield was maximum in P. florida after 12thday of fermentation. The FTIR and SEM also confirmed the chitin and chitosan. The extracted chitin and chitosan were subjected to antimicrobial activity, and it was found the extracted chitin and chitosan were active against the bacteria and fungi. Our findings suggest that the Pleurotus spp. are the potential candidate to produce eco-friendly chitin and chitosan in the development of drugs, artificial bone and raw material for the food industries in the near future.

Screening of efficient phosphate solubilizing fungi from mine soil and effect of phosphofungi on seed germination and vigour index of ground nut (arachis hypogaea L.) and green gram (vigna radiata L.)

Background: The use of chemical fertilizers to solve the problem of nutrient deficiency in soil has been associated with a number of environmental problems.Objective: The aim of this study was to perform an isolation and screening of native phosphofungi from mine soil. To evaluate the effect of phosphofungi on seed germination and seedling vigour index of Ground nut and Green gram.Materials and Methods: The phosphofungi were screened using Pikovskaya’s agar medium with tricalcium phosphate (Ca3 (PO4)2) as the phosphorus source. The soluble phosphorus, Phosphate solubilizing activity, titrable acidity, pH and fungal biomass were determined. Mineral phosphate solubilizing (MPS) activities of fungal isolates were tested in tricalcium phosphate medium.Results: Five promising phosphate solubilizing fungal species were screened. Aspergillus niger (13±1.0 mm) and Aspergillus flavus (8±0.6mm) showed the more phosphate solubilising index than compared to other fungal isolates. Analyzing the possible phosphorus released, from 3rd, 6th, 9th, and 12th day of incubation. Antagonistic effect of phosphofungi Aspergillus flavus (65±11 mm) and Aspergillus niger (60±13 mm) showed more effective against Colletotrichum sp. than compared to other fungal isolates. Aspergillus flavus (745±20µg/ml) and Penicillium spinulosum (600±20 µg/ml) showed maximum phosphate solubilizing activity on 3rd and 12th day of incubation. Fusarium redolens showed the more effectiveness on seed germination and seedling vigour than compared to other fungal isolates. Aspergillus flavus showed positive result for the production of Indole 3 acetic acid and remain four fungal strains are negative result.Conclusion: Biofertilizers from native phosphofungi could be used alongside reduced levels of inorganic fertilizers to enhance soil available Phosphorous.

Effects of Prebiotics and Synbiotics Delivered In Ovo on Broiler Small Intestine Histomorphology During the First Days After Hatching.

The objective of the study was to determine the effect of prebiotics and synbiotics administered in ovo on the 12th day of incubation, on the development of the intestinal villi and the number of goblet cells in the small intestine of broiler chickens on the Ist and the 4th days of life of chicks. Two prebiotics: inulin (PI) or Bi2tos (PB) and two synbiotics: inulin + L. lactis subsp. lactis (SI) or Bi2tos + L. lactis subsp. cremoris (SB) were injected in ovo on the 12th day of embryonic development. The control group of the embryos was injected with physiological saline (C). On the 1st day of life, an increase in the height of the villi in the jejunum was reported as a result of the injection of pre- and synbiotics, moreover an increase in the surface area of the villi in the jejunum and the duodenum in chicks from the SB group was also observed. A stimulatory effect of synbiotics on the morphology of the duodenum and thejejunum was also observed on the 4th dh day after hatching. Conversely, in the ileum, in the SB group, a reduction in the height of villi was found both on thse 1t and thte 4h days of life. In contrast, injection of inulin and synbiotic with the addition of inulin resulted in an increase in the number of goblet cells in the duodenum and the jejunum on thst 1" day of life, and caused a significant decrease on the 4th day after hatching.

In ovo exposure of Eurycoma longifolia extract on the pre-hatched development of chicken embryo

This study was conducted to investigate the effects of in ovo administration of Eurycoma longifolia (Tongkat Ali) extract (TAE) on the survivability and pre hatch development of chicken embryo. A dose response study of 0.1 mg, 1 mg and 10 mg TAE/100g egg on embryo survivability during first 5 days of incubation showed that the 0.1 to 1mg exposures had no effect on embryo survivability, and the lethal dose to kill 50% (LD50) was at around 10 mg. The 0.1 mg exposure was found to be non-lethal to the developing embryo until 19th day of incubation and it also enhanced (p<0.05) the somite development (42 in TAE and 38 in control embryos) in 88 to 96 hours embryo. Then 0.1 mg TAE exposure resulted in 70.5% body mass gain (% to egg weight) in the 19th day embryo in contrast to53.4% in the control. Compared to control, significant increases in limb lengths and feather-follicles in 9-day old embryos were observed in the 0.1 mg TAE group. On 19th day of incubation, the 100% of the 0.1mg TAE exposed embryos were found to produce traceable vocal sounds along with the pipping marks on egg shell, but these phenomena were almost absent in the control chicks during the same period of incubation. Results suggested that the TAE exposure at >1 mg/100 egg dose is lethal to developing chicken embryo, but the 0.1 mg TAE/100g egg exposure through in ovo injection can be a beneficial strategy to produce heavier and stronger day old chicks.Bang. J. Anim. Sci. 2016. 45 (2): 24-29

Partial acidulation of phosphate rock for enhanced phosphorus availability in alluvial soils of Bihar, India

The present study was undertaken to fractionate partially acidulated phosphate rock (PAPR) and evaluate its effect on Phosphorus availability in alluvial soils. For this purpose, low grade phosphate rock was collected from Udaipur, Rajasthan and acidulated at different degrees with sulphuric acid (H2SO4) (v/v)Results from an incubation study showed that the available fraction of P [Water soluble (WSP) and Citrate soluble (CSP)] increased with increasing degree of acidulation. The maximum WSP was obtained at 50% H2SO4 (52.30% of total P) and the maximum CSP was obtained at 80% H2SO4 (49.54% of total P). 50 % dissolution of rock phosphate was the best treatment and found to be at par with that of 60% and 80% but was significantly superior to rest of the treatments. The maximum WSP in the soil samples was obtained after acidulation with 50 %H2SO4 and the maximum Olsen’s P by the 14th day of incubation. An increasing trend was found at initial stages of incubation (7-14 days) with a notable decrease in the later stages. WSP demonstrated the maximum correlation at 40% dissolution(r = 96* for first order reaction, r =95* for second order reaction) while for Olsen’s P, the maximum correlation was found in initial rock phosphate sample (r =88* for first order reaction, r =94* for second order reaction). Kinetics analysis of the obtained rock phosphate demonstrated that the PARP has more influence on phosphorus release pattern as compared to the inorganic P (SSP).

Interaction effect of nitrogen and vermicompost in the presence of herbicide (Clodinafop propargyl) on nitrogen transformation in a sandy soil

A laboratory experiment with three levels of nitrogen (0, 100 and 200 mg kg-1), two levels of vermicompost (0 and 1 % on dry wt. basis) and two levels of herbicide (0 and 60 g a.i. ha-1) was conducted with sandy soil of Hisar to study the interaction of nitrogen and vermicompost in the presence of herbicide (clodinafop propargyl) on nitrogen transformation during 2014. NH4+-N contents increased upto the 14th day in soil and then declined up to 56th day under control. NO3--N content in soil increased significantly throughout the incubation study under control. With con-junctive use of nitrogen along with vermicompost, NH4+-N contents increased significantly in the soil upto 14th day of incubation with an increase from 44.49 to 73.22 mg kg-1 and 64.00 to 102.87 mg kg-1, whereas NO3--N content in soil significantly increased throughout the incubation study over control and the increase was from 13.68 to 101.36 mg kg-1 and 23.19 to 115.48 mg kg-1. However, NH4+-N and NO3--N decreased significantly at all incubation periods with the application of herbicide alone and in presence of nitrogen as well as vermicompost. The study revealed that judi-cious use of N, leads to more availability of N to crop and prevents the environmental pollution. Higher levels of N application may increase the risk of ground water pollution due to more availability of NO3- ion which can be sub-jected to leaching losses. Vermicompost proved to be the important source of nutrients as it has narrow C:N ratio and decompose more quickly than other organic manures such as FYM etc. Among commonly used herbicides, clodinafop propargyl is most commonly used herbicide and may have negative impact on the microbial population and thus may hinder the transformation processes particularly in sandy soils which have low organic matter content and thereby affecting the availability of nutrients to crop and play decisive role in crop yields.

Effect of copper nanoparticles and copper sulphate on metabolic rate and development of broiler embryos

Copper (Cu) is regularly used as a growth promoter in poultry production. However, it has been demonstrated that the content of Cu inside eggs might not be sufficient to support the embryonic development. It is possible to supply the embryo with extra nutrients by in-ovo administration. Recently, it has been shown that in-ovo administration of copper nanoparticles (Cu-NP) and copper sulphate (CuSO4) remarkably improved the body weights of growing chickens. Thus, the objective of the present experiment was to elucidate the potential effects of Cu-NP and CuSO4 on the metabolic rate (oxygen consumption — O2 and energy expenditure — EE) and development during embryogenesis.Fertilised broiler eggs were divided into six groups: a non-injected control, a placebo injected with demineralised water, two groups injected, at day one of incubation, with CuSO4 (50 and 100mg/kg) and two groups injected with Cu-NP (50 and 100mg/kg). Gaseous exchange was measured in an open-air-circuit respiration unit, and EE was estimated from day 10 to day 19 of embryogenesis. Body weight at 24h after hatching and the relative organ weights were used as a measure of hatching development. In-ovo injection of 50mg/kg Cu-NP and CuSO4 significantly increased O2 consumption and EE on the 16th and 19th day of incubation compared with the control group; Cu-NP had the largest effect on the metabolic rate. However, organ weights (intestine, heart, liver, and breast) relative to the yolk-free body weight were not affected in the injected groups. In addition, blood parameters did not show any changes among the groups. This result demonstrates that in-ovo injection of Cu-NP affects the metabolic rate of embryos, which might explain their improved performance after hatching.

Gamma radiation inhibits the production of Ochratoxin A by Aspergillus carbonarius. Development of a method for OTA determination in raisins

Ochratoxin A (OTA) is a mycotoxin of great interest to humans for its nephrotoxic effects. The development and validation of an OTA method determination in raisins is described by using High Pressure Liquid Chromatography (HPLC) with Fluorescence Detector (FD). The recovery of the method was 104% while the detection limit (DL) and quantification limit were 0.26 and 0.51 ng g−1, respectively. The effect of gamma irradiation at dose of 10 kGy on the production of OTA by Aspergillus carbonarius in raisins (Vitis vinifera L.) and on OTA in contaminated samples, was investigated. The OTA which was produced by A. carbonarius on the 12th day of incubation, after irradiation, showed that at dose level of 10 kGy, OTA production was not detectable (below the detection limit of the method) compared with the non-irradiated the same day. A dose of 10-kGy gamma radiation applied on 100 ng of OTA, in raisins reduced OTA contamination ~88%. According to the risk assessment analysis the Provisional Maximum Tolerable Daily Intake (PMTDI) of 5.8 ng OTA kg−1 bw, indicates that adult consumers are less exposed, 31.4–78.4 fold lower to OTA by the irradiated raisins.

Neurotoxicological effects of nicotine on the embryonic development of cerebellar cortex of chick embryo during various stages of incubation.

Long-acting nicotine is known to exert pathological effects on almost all tissues including the cerebellar cortex. The present work was designed to elucidate the effect of nicotine on the development of cerebellar cortex of chick embryo during incubation period. The fertilized eggs of hen (Gallus gallus domesticus) were injected into the air space by a single dose of long acting nicotine (1.6 mg/kg/egg) at the 4th day of incubation. The embryos were taken out of the eggs on days 8, 12 and 16 of incubation. The cerebellum of the control and treated embryos at above ages were processed for histopathological examination. The TEM were examined at 16th day of incubation. The results of the present study revealed that, exposure to long-acting nicotine markedly influence the histogenesis of cerebellar cortex of chick embryo during the incubation period. At 8th day of incubation, nicotine delayed the differentiation of the cerebellar analge; especially the external granular layer (EGL) and inner cortical layer (ICL). Furthermore, at 12th day of incubation, the cerebellar foliation was irregular and the Purkinje cells not recognized. By 16th day of incubation, the cerebellar foliations were irregular with interrupted cerebellar cortex and irregular arrangement of Purkinje cells. Immunohistochemical analysis for antibody P53 protein revealed that the cerebellar cortex in all stages of nicotine treated groups possessed a moderate to weak reaction for P53 protein however; this reaction was markedly stronger in the cerebellar cortex of control groups. Moreover, the flow cytometric analysis confirmed that the percentage of apoptosis in control group was significantly higher compared with that of nicotine treated group. At the TEM level, the cerebellar Purkinje cells of 16th day of treated groups showed multiple subcellular alterations in compared with those of the corresponding control group. Such changes represented by appearing of vacuolated mitochondria, cisternal fragmentation of RER, irregular grooves of Golgi tubules. Also, multiple cytoplasmic vacuoles and aggregation of Nissl granules were recorded around pyknotic nucleus.

Study on Teratogenic Effects of Bhavana Panjankula Thailam, Castor oil, Ricin Using Chick Embryo as a Model System

Teratogenicity is the ability to cause developmental abnormaltiies in foetus. Substances with teratogenicity effects can damage the DNA of a developing foetus. They may cause abnormal development of a limb malformation of an organ and the effects for the developing foetus can vary depending on the teratogen. Chicken eggs are used as a model system to test teratogenic effect of drug. The present study carried out, Bhavana panjankula thailam is one of the Siddha drug indicated for karuppachudu (a type of pitha disorder affecting uterus), urinary tract infections, dry skin and constipation. In this study five concentrations of drug Bhavana panjankula thailam were injected into the chicken embryos. The castor oil, Ricin also injected to chick embryos at same concentrations. Lithium is used as a positive control in the same concentrations which affects limb deformities of the organ. 1: 1 ratio of lithium and drug were injected into the embryo for scrutinize the effect of drug. At the 12th day of incubation the eggs were dissected out and detect the malformation in the embryo. The results suggested that the lowest concentrations of drug treated embryo have no malformation and highest concentrations of drug treated embryo have malformed like heart and other organs are distorted. The DNA was isolated from embryos of chick and run in agarose gel electrophoresis to confirm the DNA damage.

Proliferative and oxidative response of hepatocytes (Hep) and hepatic stellate cells (HSC) isolated from rats exposed to ketogenic diet.

Ketogenic diet (KD) is considered in the context of its anti-epileptic effects, but its influence on liver dysfunction has not been elucidated yet. The study was aimed to investigate the activity of hepatocytes (Hep) and hepatic stellate cells (HSC) isolated from rats fed with KD, in respect of NO and superoxide generation by these cells as well as their proliferative activity in vitro. We also sought to characterize the plasma FFA profiles in control and ketogenic rats. Hep and HSC were isolated by the collagenase perfusion method and separated by the Percoll gradient centrifugation. After the 4th, 8th and 12th day of incubation, the media were collected for further analysis. NO generation increased within the time of incubation both in Hep and HSC isolated from KD-rats. In HSC group NO production raised significantly from 2.65 ± 0.07 μM/10(6) cells on 4th day of incubation to 5.49 ± 1.2 μM/10(6) cells on 12th day of incubation. In respect to O2⁻· generation experimental Hep and HSC provide considerably higher quantities of this free radical until 12th day of incubation (2.5 ± 0.07 and 3.2 ± 0.3 nM/10(6) cells, respectively). Although KD exerts anti-proliferative effect on hepatocytes, in respect to HSC it intensifies their proliferative activity. Furthermore, as we estimated on the basis of NO and O2⁻. generation both Hep and HSC exposed to KD are the source of free radicals.