Immunological effect of Moringa Oleifera leaf extract on vaccinated and non-vaccinated Hubbard chickens experimentally infected with Newcastle virus

In veterinary medicine plant based medicine is achieving a huge importance worldwide. This research was subjected to rectify the hydrophilic Moringa Oleifera alcoholic leaves extract could improve the immune system in vaccinated and non-vaccinated broiler Hubbard chickens experimentally exposed to Newcastle disease (ND) virus. Seventy five chicks with age one day old were splitted randomly into five groups equally in distribution with fifteen chick in each group. Group I was untreated unvaccinated (control negative group) while group IV was infected group with NDV (control positive group). The experimental Groups II and V were given daily oral treatment of hydrophilic alcoholic leaves extract of M. oleifera at 200 mg/kg body weight until day 21 of age while groups III and V were ND vaccinated with La Sota strain of ND vaccines. The four groups (II, III, IV, V) were infected with ND virus velogenic strain (VNDV) on day 21. Following to infection, Monitoring of birds were done daily for clinical signs, postmortem examination, morbidity and mortality. Cellular, humeral immune response and phagocytic activity were evaluated and the data were statistically analyzed using (SPSS). Total and differential cell numbers as well as Haemagglutination inhibition (HI) titre increased in the extract treated and vaccinated group which give total protection against NDV much more than treated and unvaccinated group. As a result it could be recommended to use M. Olifera extract from the first day of rearing in Hubbard chicken with ND vaccination program as a prophylactic treatment in protection of birds against ND infection.     

Lactobacilli and Bifidobacteria Promote Immune Homeostasis by Modulating Innate Immune Responses to Human Rotavirus in Neonatal Gnotobiotic Pigs

The effects of co-colonization with Lactobacillus rhamnosus GG (LGG) and Bifidobacterium lactis Bb12 (Bb12) on 3-dose vaccination with attenuated HRV and challenge with virulent human rotavirus (VirHRV) were assessed in 4 groups of gnotobiotic (Gn) pigs: Pro+Vac (probiotic-colonized/vaccinated), Vac (vaccinated), Pro (probiotic-colonized, non-vaccinated) and Control (non-colonized, non-vaccinated). Subsets of pigs were euthanized pre- [post-challenge day (PCD) 0] and post (PCD7)-VirHRV challenge to assess diarrhea, fecal HRV shedding and dendritic cell/innate immune responses. Post-challenge, Pro+Vac and Vac groups were completely protected from diarrhea; protection rates against HRV shedding were 100% and 83%, respectively. Diarrhea and HRV shedding were reduced in Pro compared to Control pigs following VirHRV challenge. Diarrhea scores and virus shedding were significantly higher in Controls, compared to all other groups, coincident with significantly higher serum interferon-alpha levels post-challenge. LGG+Bb12 colonization ±vaccine promoted immunomaturation as reflected by increased frequencies of CD4, SWC3a, CD11R1, MHCII expressing mononuclear cells (MNCs) and conventional dendritic cells in intestinal tissues and blood post-challenge. Colonization decreased frequencies of toll-like receptors (TLR) 2 and TLR4 expressing MNCs from vaccinated pigs (Pro+Vac) pre-challenge and increased frequencies of TLR3 expressing MNCs from Pro pigs post-challenge, suggesting that probiotics likely exert anti-inflammatory (TLR2 and 4 down-regulation) and antiviral (TLR3 up-regulation by HRV dsRNA) actions via TLR signaling. Probiotic colonization alone (Pro) increased frequencies of intestinal and systemic apoptotic MNCs pre-challenge, thereby regulating immune hyperreactivity and tolerance. However, these frequencies were decreased in intestinal and systemic tissues post-challenge, moderating HRV-induced apoptosis. Additionally, post-challenge, Pro+Vac and Pro groups had significantly decreased MNC proliferation, suggesting that probiotics control excessive lymphoproliferative reactions upon VirHRV challenge. We conclude that in the neonatal Gn pig disease model, selected probiotics contribute to immunomaturation, regulate immune homeostasis and modulate vaccine and virulent HRV effects, thereby moderating HRV diarrhea.     

Sequence heterogeneity of the ribosomal DNA inAllium cepa (Liliaceae)

Summary Southern blot hybridizations were performed to investigate the ribosomal DNA structure inAllium cepa (Liliaceae). The analysis of single and double digestions with Eco RI, Hind III, and Bam HI endonuclease restriction enzymes showed the presence of at least two types of repetitive units. The gene types were present in each single individual ofAllium cepa analyzed, notwithstanding the variation of the ribosomal gene number from 5,500 to 11,900 and the NOR number from 2 to 4. The first and the second gene type are 12.7 kb long and the first type is much more represented inAllium cepa genome. The differences between the two gene types consist in the position of the Hind III restriction sites in the external spacer.Work supported by CNR contract no. 74/0267.     

Allium cepa assay based comparative study of selected vegetables and the chromosomal aberrations due to heavy metal accumulation

Irrigation of industrial effluents may end in the bioaccumulation of various toxic metals and consequent genetic changes in contaminated food crops. To test this hypothesis and extent of genetic modifications, Allium cepa test was performed to food crops viz. tomato (Lycopersicum esculentum) and chili (Capsicum annum) as Allium cepa test is a useful tool to assess genetic variations in plants. Prior to A. cepa test, the plants were exposed to various metal concentrations 125–1000 mg/L in the synthetic wastewater. The extracts of harvested plants were used to grow the root of A. cepa following its standard method. The root tips were fixed, stained and examined under compound microscope (almost 300–400 dividing cells) to check the extent of chromosomal variations during various stages of mitosis. The results revealed various chromosomal abnormalities including laggards, stickiness, vagrant chromosomes, binucleated cells, nuclear lesions, giant cells and c-mitosis at different level of treatment. On the whole, aberrations were increasing with the increasing doses along the positive control. In comparison, chili crop had higher level of aberrations depicting the higher chromosomal changes. Lower mitotic index (MI) with increasing level of doses was also describing the hampered cell division due to increased metal stress. The study is showing that the cell division was ceased with increasing metal stress thus increasing the rate of cell aberrations.     

Evaluation of cefquinome's efficacy in controlling avian colibacillosis and detection of its residues using high performance liquid chromatography (HPLC)

This study aimed to evaluate the efficacy of cefquinome in treatment and controlling of Escherichia coli experimentally infected broiler chickens, in addition of detection of its residues using High performance liquid chromatography (HPLC). In this study, 150 one-day old Cobb broiler chicks were used. On the 14^th day chicks experimentally infected and divided into 3 equal groups (50 each); control group (G1) non-infected, non-treated, (G2) infected with E. coli O₇₈ non treated, (G3) infected with E. coli O₇₈, cefquinome treated. Cefquinome was administrated 5^th day post infection, intramuscularly by a dose of (2 mg/ kg b w.t) for 3 consecutive days. Experimental E. coli infection in broilers induced weakness, loss of appetite, depression, cough and watery diarrhea in addition to a recorded mortality (30%) with reduction in growth performance, erythrogram, total proteins, albumin, antioxidants and haemagglutination inhibition (HI) titers. In addition, a significant increase in feed conversion rate (FCR), leukocytic count, liver enzymes, kidney functions, total globulins, malondialdehyde, nitric oxide and lysozyme activity. Treatment with cefquinome led to decreased mortality rate, improvement in clinical signs, growth performance and modulated most of these altered parameters. Cefquinome's residues was not detected in breast muscles 3^rd day and liver and kidneys 7^th days post treatment. Therefore, it's recommended that cefquinome is a good choice for controlling of colibacillosis in broilers and its withdrawal time 3 days in breast muscles and 7 days in liver and kidney post treatment.     

Eimeria tenella: utilization of a nasal vaccine with sporozoite antigens incorporated into Iscom as protection for broiler breeders against a homologous challenge

The aim of this study was to evaluate a nasal vaccine using antigens derived from sporozoites of Eimeria tenella incorporated into Iscom to protect broiler chicks. Forty-five one-day-old chickens (Cobb), unvaccinated against coccidiosis, were used in this experiment. The birds were maintained in separated battery cages and divided into three groups: G1 (n = 15), G2 (n = 15), and G3 (n = 15). G1 received 50 μg of sporozoites + Iscom vaccine, G2 received Iscom without antigens, and G3 received only PBS. The treatments were administered by nasal route on days 0, 7, and 21 of the experiment. On the 28th day, all birds were challenged with 105 sporulated oocysts of E. tenella. On the challenge day, three birds from each group were euthanized to evaluate lymphocyte proliferation. Lesion scores were obtained from five birds from each group, 7 days after challenge. The remaining animals were euthanized on the 50th day. The mean lymphocyte proliferation responses were significantly different (P = 0.03); G1 was 2.3-2.6 times more elevated than G2, and G3 (P < 0.001). 83% of the birds from G1 showed an IgY antibody reaction by ELISA at challenge. The means for oocysts shedding were 16,890 ± 20,511, 48,080 ± 50,047, and 65,020 ± 74,461, for G1, G2 and G3 birds, respectively. There was no significant difference (P > 0.17) in oocysts shedding between groups. However, the G1 and G2 chicks demonstrated reduction in percentage of oocyst shedding when compared to control birds (G3) by 74.02% and 26.05%, respectively. The average lesion scores were G1 = 0.4, G2 = 1, and G3 = 2. This study demonstrated that the lowest lesion score and oocyst shedding were observed in the birds from the group that received antigens derived from sporozoite with an Iscom adjuvant (G1). These results suggest that this vaccine can induce protection against avian coccidiosis.     

Immune Response in Goats to Different Payloads of FMDV Monovalent Vaccine: Protection Against Virulent Challenge and Development of Carrier Status

The relationship of Foot-and-Mouth Disease virus antigen payload and number of dose of vaccine conferring protection against virus challenge in goats was studied. Goats vaccinated with oil adjuvant Foot-and-Mouth Disease vaccines containing different antigen payloads with or without booster resisted virulent challenge at 21 days post-vaccination or 7 days after booster respectively. However, localized sub-clinical infection was observed in two vaccinated goats on 35 days post-challenge. RNA could be detected from 31.8% of vaccinated goats (10^2.69–10^4.99 viral RNA copies per cotton swab of nasal secretions) on day 35 post-challenge. Since no live virus could be isolated after 5 days post-challenge, the risk of these animals transmitting the disease was probably very low. The finding showed that oil adjuvant Foot-and-Mouth Disease vaccines containing antigen payload of 1.88 μg may prevent or reduce the local virus replication at the oropharynx and shedding of virus from nasal secretions and thereby reduce the amount of virus released into the environment subsequent to exposure to live virus. This study also showed that goats with poor sero conversion to vaccination can be infected without overt clinical signs and became carriers like sheep.     

Comparison of cytotoxic and genotoxic effects of plutonium-239 alpha particles and mobile phone GSM 900 radiation in the Allium cepa test

The goal of this study was to compare the cytotoxic and genotoxic effects of plutonium-239 alpha particles and GSM 900 modulated mobile phone (model Sony Ericsson K550i) radiation in the Allium cepa test. Three groups of bulbs were exposed to mobile phone radiation during 0 (sham), 3 and 9 h. A positive control group was treated during 20 min with plutonium-239 alpha-radiation. Mitotic abnormalities, chromosome aberrations, micronuclei and mitotic index were analyzed. Exposure to alpha-radiation from plutonium-239 and exposure to modulated radiation from mobile phone during 3 and 9 h significantly increased the mitotic index. GSM 900 mobile phone radiation as well as alpha-radiation from plutonium-239 induced both clastogenic and aneugenic effects. However, the aneugenic activity of mobile phone radiation was more pronounced. After 9 h of exposure to mobile phone radiation, polyploid cells, three-groups metaphases, amitoses and some unspecified abnormalities were detected, which were not registered in the other experimental groups. Importantly, GSM 900 mobile phone radiation increased the mitotic index, the frequency of mitotic and chromosome abnormalities, and the micronucleus frequency in a time-dependent manner. Due to its sensitivity, the A. cepa test can be recommended as a useful cytogenetic assay to assess cytotoxic and genotoxic effects of radiofrequency electromagnetic fields.     

Influence of Glomus fasciculatum and Meloidogyne hapla on Allium cepa in Organic Soils

The influence of Meloidogyne hapla and Glomus fasciculatum on Allium cepa (onion) grown in organic soil was evaluated under greenhouse conditions. In the absence of G. fasciculatum, M. hapla significantly retarded the growth of A. cepa cv. Krummery Special and MSU 8155 × 826, but had no detrimental influence on Downing Yellow Globe, Spartan Banner, or Spartan Sleeper. All five cultivars maintained populations of M. hapla, Final root population densities of M. hapla associated with Spartan Banner, Krummery Special, MSU 8155 × 826, and Spartan Sleeper were significantly greater than those recovered from Downing Yellow Globe. Final root population densities of M. hapla were directly proportional to the initial population densities. Root colonization of onion by G. fasciculatum significantly enhanced the growth and development of Downing Yellow Globe. The rate of increase of A. cepa growth and the final spore density were directly proportional to the initial spore density of G. fasciculatum. Final population densities of M. hapla in the presence of G. fasciculatum were generally greater than in the absence of the fungus. After 15 weeks, A. cepa plants grown in the presence of both M. hapla and G. fasciculatum were significantly larger than those grown in the presence of only M. hapla.     

Influence of Glomus fasciculatum on Meloidogyne hapla Infecting Allium cepa

The impact of Glomus fasciculatum on Meloidogyne hapla associated with Allium cepa was evaluated in two experiments. Nematode density was not different in mycorrhizal and nonmycorrhizal plants 10 weeks after the joint inoculation of M. hapla and G. fasciculatum. Differences in the age structure of M. hapla populations reared on mycorrhizal and nonmycorrhizat plants were noted. G. fasciculatum enhanced leaf and bulb growth of A. cepa in the absence of M. hapla, but did not affect plant weight when nematodes were present. Survival and reproduction of M. hapla were not affected by G. fasciculatum or phosphorus (P). The estimated time required for inoculated second-stage juveniles (J2) to mature to the adult stage was 1,000 degree hours (base = 9 C) greater in mycorrhizal than in nonmycorrhizal plants supplemented with P. Although the infectivity of J2 was not measured directly, colonization of A. cepa by G. fasciculatum appeared to alter the ability of M. hapla to penetrate roots.     

Biochemical degradation pathway of textile dye Remazol red and subsequent toxicological evaluation by cytotoxicity, genotoxicity and oxidative stress studies

Remazol red (RR), a monochloro sulphonated azo dye was degraded up to 97% within 20 min at 40 °C and pH 7 at dye concentration 50 mg l⁻¹ by Pseudomonas aeruginosa BCH. Examination of enzyme status exposed the involvement of various oxidoreductive enzymes viz. laccase, veratryl alcohol oxidase and NADH-DCIP reductase. Analytical studies viz. HPTLC, HPLC, FTIR and GC-MS carried out with dye and dye metabolites formed after dye decolorization confirmed that the decolorization was due to degradation. Based on enzymatic status and GC-MS analysis the possible metabolic pathway followed by bacterial strain for the degradation of RR was proposed. During toxicological scrutiny, cell death was observed in RR treated Allium cepa (A. cepa) root cells. The observed inhibition of catalase (CAT) activity and induction in enzyme activities of sulfur oxide dismutase (SOD) and ascorbate peroxidase (APX) along with raised protein oxidation and lipid peroxidation signified that RR generated the oxidative stress in A. cepa roots. These toxicological studies along with genotoxicity studies using A. cepa roots and phytotoxicity studies using Phaseolus mungo (P. mungo) and Sorghum vulgare (S. vulgare) conclusively designated the toxicity of RR and comparatively less toxic nature of metabolites formed after dye degradation by P. aeruginosa BCH.     

Synthesis of eco-friendly silver nanoparticles using Allium sp. and their antimicrobial potential on selected vaginal bacteria

Allium cepa and garlic Allium sativa plants were used to evaluate their potential synthesis of silver nanoparticles and their antibacterial effect on Streptococcus pneumoniae and Pseudomonas aeruginosa. Transmission electron microscopy (SEM) was used to distinguish the morphology of the nanoparticles attained from plant extracts. Energy dispersive X-ray (EDX) spectrometer established the existence of elemental sign of the silver and homogenous allocation of silver nanoparticles. Diffraction by using X ray (XRD) analysis for the formed AgNPs revealed spherical plus cubical shapes structure with different planes ranged between 111 and 311 planes. The antibacterial action of AgNPs against vaginal pathogens, Streptococcus pneumoniae and Pseudomonas aeruginosa was recognized. Our work showed a rapid, eco-safety and suitable method for the synthesis of AgNPs from Allium cepa and garlic Allium sativa extracts and can be used in biomedical applications.     

Genotoxicity of five food preservatives tested on root tips of Allium cepa L.

The effects of the food preservatives sodium benzoate (SB), boric acid (BA), citric acid (CA), potassium citrate (PC) and sodium citrate (SC) have been studied on root tips of Allium cepa L. Roots of A. cepa were treated with a series of concentrations, ranging from 20 to 100 ppm for 5, 10 and 20 h. The results indicate that these food preservatives reduced mitotic division in A. cepa compared with the respective control. Mitotic index values were generally decreased with increasing concentrations and longer treatment times. Additionally, variations in the percentage of mitotic stages were observed. The total percentage of aberrations generally increased with increasing concentrations of these chemicals and the longer period of treatment. Different abnormal mitotic figures were observed in all mitotic phases. Among these abnormalities were anaphase bridges, C-mitosis, micronuclei, lagging, stickiness, breaks and unequal distiribution.     

Evaluation of malathion-induced cytogenetical effects and oxidative stress in plants using Allium test

The present study emphasized to explore the toxicity effect of malathion on plants using Allium test. The experiments explored the mitotic inhibition, growth and activity of antioxidant enzymes in roots of Allium cepa at different concentrations (50, 125, 250 and 375 ppm) of malathion under different exposure periods (3, 9 and 18 h). The results revealed that all concentrations of malathion were capable to decline the root growth. Malathion-induced mitotic alterations varying from reduction in mitotic index (MI), relative division rate (RDR) and phase distribution along with large number of chromosomal aberrations. These changes were of varying degree depending on the concentration and treatment period. The roots treated with malathion (375 ppm) showed significant (p ≤ 0.05) higher levels of superoxide dismutase and catalase than the control, while the activity of peroxidase was significantly (p ≤ 0.05) low. At 375 ppm malathion, malondialdehyde content was significantly high (p ≤ 0.05) that was increased with the treatment period. Findings concluded that variations in mitotic index, chromosomal aberrations, alterations in malondialdehyde content and activities of antioxidant enzyme could serve as the useful indicators for monitoring the effects of malathion exposures in the real scenario. Superoxide dismutase and catalase enzyme play vital roles in the antioxidant defence mechanisms under malathion toxicity. According to the data on the malondialdehyde content show malathion to be capable of producing superoxide radicals indirectly, and to result in membrane damage and oxidative stress.     

Heterologous expression,characterization and evaluation of the matrix protein from Newcastle disease virus as a target for antiviral therapies

Newcastle disease virus (NDV) is an infectious agent of a large variety of birds, including chicken, which poses a real threat to the agriculture industry. Matrix (M) proteins of NDV and many other viruses perform critical functions during viral assembly and budding from the host cell. M-proteins are well conserved and therefore are potential targets for antiviral therapies. To validate this, we expressed the NDV M-protein in its native form in Saccharomyces cerevisiae and in inclusion bodies in Escherichia coli. Proper refolding of the recombinant protein produced in E. coli was verified using circular dichroism and infrared spectroscopies and electron microscopy. Immunization of chickens with the NDV M-protein elicited significant serum antibody titers. However, the antibodies conferred little protection against the ND following lethal viral challenges. We conclude that the M-protein is not exposed on the surface of the host cell or the virus at any stage during its life cycle. We discuss how the conserved M-protein can further be exploited as an antiviral drug target.     

Protein synthesis requirements at specific points in the interphase of meristematic cells

A 6 h treatment with anisomycin at a concentration of 1 μg/ml enables us to modify the steady-state kinetics of a meristematic cell population of Allium cepa, and this points to a difference of sensitivity to inhibition of protein synthesis between the several periods of the cell division cycle (G1, S, G2, M). The results show that the cells are incapable of entering the S period in the presence of the inhibitor, but that, where DNA synthesis has already been initiated, the synthesis continues in the cells in question. It was found, moreover, that there is a point in the early G2 period, which has a duration of approx. 3 % of the total duration of the cycle, at which the synthesis of specific proteins appears to determine the progression of cells to mitosis.     

Effects of aluminum on nucleoli in root tip cells and selected physiological and biochemical characters in <Emphasis Type="Italic">Allium cepa</Emphasis> var. <Emphasis Type="Italic">agrogarum</Emphasis>L

Background  

Increased Al concentration causes reduction of mitotic activity, induction of nucleolar alteration, increase of the production of ROS and alteration of several antioxidant enzyme activities in plant cells. Allium cepa is an excellent plant and a useful biomarker for environmental monitoring. Limited information is available about the effects of Al on nucleoli, antioxidant enzyme system, contents of MDA and soluble protein in A. cepa. Therefore, we carried out the investigation in order to better understand the effects of Al on the growth, nucleoli in root tip cells and selected physiological and biochemical characters.     

Irreversible effects of ionizing RTG-radiation on the osmotic properties of the epidermal cells ofAllium cepa L. bulbs

The effect of different exposures to RTG radiation (100 r, 250 r, 500 r, 1000 r, 2500 r, 5000 r) on the changes of the permeability of the epidermal cells ofAllium cepa L., variety V?etaty, bulbs was studied during three vegetation periods. Bulbs from growing plants (followed from the development phase of bulb growth) and bulbs immediately after harvesting were used for the experiments. It was demonstrated in our experiments by plasmolysis that the radiation-induced changes of cell permeability were irreversible. When plasmolytic agents of non-electrolyte character (1M sucrose, 1M glycerol, 1M urea) were used, the equilibrium state (total plasmolysis) was reached more rapidly with the irradiated bulbs than with the unirradiated controls. When electrolytes were used (0·5M KNO₃, 0,5M KBr), total plasmolysis occurred less rapidly with the irradiated objects than with the controls. These relationships were manifest during the whole ontogenesis ofAllium cepa L. An exposure to 100 r was sufficient in our experiments for causing the mentioned changes, proving a considerable sensitivity of the osmotic properties of the epidermal cells ofAllium cepa L. bulbs to ionizing RTG radiation. Plasmolytic experiments after smaller exposures than 100 r of RTG-rays are in preparation.     

Cytogenetic effects of silver and gold nanoparticles on Allium cepa roots

The present study evaluates the cytogenetic effects of both silver and gold nanoparticles on the root cells of Allium cepa. In this study, the root cells of Allium cepa were treated with both gold and silver nanoparticles of different concentrations (1?mg/L, 5?mg/L and 10?mg/L) along with control for 72?h. Experimental results revealed that after 72?h of exposure, a significant decrease in mitotic index (MI) from 68% (control) to 52.4% (1?mg/L), 47.3% (5?mg/L) and 41.4% (10?mg/L) for gold nanoparticles and 57.1% (1?mg/L), 53% (5?mg/l), 55.8% (10?mg/L) for silver nanoparticles. Through minute observation of the photograph, it was recorded that some specific chromosomal abnormalities such as stickiness of chromosome, chromosome breaks, nuclear notch, and clumped chromosome at different exposure conditions. Therefore, present results clearly suggest that Allium cepa root tip assay could be a viable path through which negative impact of both gold and silver nanoparticles can be demonstrated over a wide range of concentrations.