Yersinia ruckeri,the causative agent of enteric redmouth disease in fish

Enteric redmouth disease (ERM) is a serious septicemic bacterial disease of salmonid fish species. It is caused by Yersinia ruckeri, a Gram-negative rod-shaped enterobacterium. It has a wide host range, broad geographical distribution, and causes significant economic losses in the fish aquaculture industry. The disease gets its name from the subcutaneous hemorrhages, it can cause at the corners of the mouth and in gums and tongue. Other clinical signs include exophthalmia, darkening of the skin, splenomegaly and inflammation of the lower intestine with accumulation of thick yellow fluid. The bacterium enters the fish via the secondary gill lamellae and from there it spreads to the blood and internal organs. Y. ruckeri can be detected by conventional biochemical, serological and molecular methods. Its genome is 3.7 Mb with 3406–3530 coding sequences. Several important virulence factors of Y. ruckeri have been discovered, including haemolyin YhlA and metalloprotease Yrp1. Both non-specific and specific immune responses of fish during the course of Y. ruckeri infection have been well characterized. Several methods of vaccination have been developed for controlling both biotype 1 and biotype 2 Y. ruckeri strains in fish. This review summarizes the current state of knowledge regarding enteric redmouth disease and Y. ruckeri: diagnosis, genome, virulence factors, interaction with the host immune responses, and the development of vaccines against this pathogen.     

A methodological study on participatory barley breeding II. Response to selection

Farmer participation is increasingly seen as a key to develop technologies which are more relevant to farmers' communities. In plant breeding, farmer participation is seen as key to increase the probability of adoption of new varieties. This paper addresses the issue of selection efficiency in participatory plant breeding by testing the effect of selection environment and of who did the selection in one cropping season (1997) on the performance of the selected lines in the following cropping season (1998). Selection environment had a larger effect on response to selection than who did the selection, confirming the importance of decentralized selection. Selections made by the breeder and the farmers in 1997, differed in 1998 for a number of traits, but seldom for grain yield. When the difference for grain yield was significant, breeder's selection was more effective on station, while farmers' selection was more effective in farmers' fields. The results of this study indicate that it is possible to organize a plant breeding program with the objective of adapting crops to a multitude of both physical and socio-economic environments: such a breeding program will, at the same time, increase productivity and stability, enhance biodiversity and produce environmentally friendly cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Prevalence pattern and biology of Sarcocystis capracanis infection in the Egyptian goats: a light and ultrastructural study

Cysts of Sarcocystis capracanis obtained from infected goats were examined to clarify the effect of the parasite on the host. Muscle tissues from fresh oesophagus, tongue, diaphragm and skeletal muscles of 680 goats were slaughtered in the main abattoir of Cairo, Egypt and they were examined microscopically for Sarcocystis infection for the first time in Egypt. 540 out of 680 (79.4%) of examined goats were found to be infected with Sarcocystis sp. The infection was recorded firstly by light microscopy as spindle shaped cysts embedded in the muscle tissues. The validity of this species as S. capracanis was confirmed by means of ultrastructural characteristics of the primary cyst wall which revealed the presence of thick-radially striated wall with finger like projections, underlined by a thick layer of ground substance enclosing the developing metrocytes and merozoites that usually contain nearly all the structures of the apical complex and fill the interior cavity of the cyst. The cyst cavity is divided by many septa extending from the ground substance and producing large number of chambers. An experimental infection using the highly infected muscles was carried out to determine the final host, which is dog. Smears of intestinal epithelium were taken to examine the endogenous stages (gamogony and sporogony) by means of light microscopy. These stages were mainly observed as to infect the lamina propria of the posterior third of the small intestine. Gamogony and zygote formation (fertilization) occurred 2-8 days post infection, while sporulation took place within the final host 13-15 days and sporocysts were passed within faeces of the infected puppies at that time. The prepatent period of S. capracanis was 12-15 days, while the patent period was extended to 37 days. In goats, infection with S. capracanis led to the loss of weight, anaemia, abortion and even death in cases of heavy infection. While bleeding, watery faeces filled with mucous on 5th and 8th day p.i. as well as intestinal lesions are the pathogenic effects occurred in puppies after experimental infection.     

Synergistic Effect of Biosynthesized Silver Nanoparticles and Natural Phenolic Compounds against Drug-Resistant Fish Pathogens and Their Cytotoxicity: An In Vitro Study

Fish pathogens causing disease outbreaks represent a major threat to aquaculture industry and food security. The aim of the presented study is to develop safe and effective bioactive agents against two bacterial isolates: Aeromonas hydrophila and Pseudomonas fluorescens. We employed a broth microdilution method to investigate the antibacterial effect of biosynthesized silver nanoparticles (AgNPs); rutin, a natural flavonoid extracted from Ruta graveneoles; and heliomycin, a secondary metabolite produced by marine actinomycetes AB5, as monotherapeutic agents. Moreover, AgNPs in combination with rutin (AgNP + R) and heliomycin (AgNPs + H) were examined for their synergistic effect. The cytotoxic effect of individual bioactive compounds and in combination with AgNPs was investigated on epithelioma papulosum cyprini (EPC) fish cell lines. Individual treatment of AgNPs, rutin, and heliomycin exhibited a dose-dependent antimicrobial activity against A. hydrophila and P. fluorescens. Rutin minimum inhibitory concentration (MIC) showed the lowest cytotoxicity when tested on EPC cell lines, while heliomycin MIC was highly cytotoxic. Combined subtherapeutic doses of AgNPs + R and AgNPs + H displayed additive and synergistic effects against A. hydrophila and P. fluorescens, respectively, with improved results and relative safety profile. The study findings demonstrate that a combination of AgNPs and natural bioactive compounds may represent novel therapeutics fighting fish pathogens potentially affecting the fish farming industry.     

Ovicidal and oviposition deterrent activities of the flavonoid pectolinaringenin from Clerodendrum phlomidis against Earias vittella

Fractions derived from Clerodendrum phlomidis Linn. F. and isolated compounds from chloroform extract were evaluated against Earias vittella Fab. for their ovicidal and oviposition deterrent activities. There were 12 fractions obtained with silica gel column chromatographic techniques using different solvent systems. Among the fractions, fraction 5 recorded maximum ovicidal activity of 68.49% and oviposition deterrent activity of 100% at 1000 ppm concentration followed by 500 ppm concentration. The active fraction was further purified using HPLC. The purified fraction was subjected to UV, ¹HNMR, ¹³CNMR, IR and EI-Mass spectroscopy and their spectral data confirmed the compound pectolinaringenin which belongs to the flavonoids. The compound pectolinaringenin was screened for its ovicidal and oviposition deterrent activities. It showed maximum ovicidal activity of 64.29% and oviposition deterrent activity of 100% at 100 ppm concentration against E. vittella. To the best of our knowledge, this is the first report of fractions and isolated compounds from C. phlomidis evaluated against E. vittella for their ovicidal and oviposition deterrent activities. The results from the present investigation clearly suggest that the active fraction or the isolated compound could be used to develop a novel pesticidal formulation to control economically important agricultural pests.     

Removal of Bisphenol A and 17β-Estradiol by Single-Walled Carbon Nanotubes in Aqueous Solution: Adsorption and Molecular Modeling

Single-walled carbon nanotubes, both before (SWNTs) and after treatment (t-SWNTs) with acidified ammonium persulfate, were successfully used to adsorb bisphenol A (BPA) and 17??-estradiol (E2) from aqueous systems. The surface characteristics of the SWNTs and t-SWNTs were analyzed by measuring their surface charge and by imaging their morphological properties through transmission electron microscopy. The extent of defects on the SWNT scaffold generated through acid etching was analyzed by Raman spectroscopy. A total of 19.4, 15.4, and 14.3?mg/g of BPA was adsorbed on SWNTs, while a total of 8.0, 6.4, and 5.1?mg/g was adsorbed on t-SWNTs with a 72-h contact time at 280, 295, and 315?K, respectively. A significantly high fraction of E2 (27.2?mg/g) was absorbed by both SWNTs and t-SWNTs, as compared to BPA. The adsorption kinetics was analyzed using a pseudo-second-order model. Sorption experiments showed that t-SWNTs adsorbed less than half as much BPA as SWNTs, but their E2 adsorption was similar. The sorption mechanism was investigated by performing molecular-level calculations. Adsorption energies calculated using density functional theory show preferential sorption of E2 to SWNTs and graphene (?26.2?kcal/mol on SWNT and ?34.1?kcal/mol on graphene) compared to BPA (?17.1?kcal/mol on SWNT and ?22.5?kcal/mol on graphene), which were consistent with the experimental findings. Thus, ab initio calculations can mechanistically explain the adsorption differences of BPA and E2 on SWNTs.     

IRMS as a tool to obtain the carbon turnover (δ13C) in organs of weaned piglets fed glutamic acid and nucleotides

Early weaning of piglets causes stress characterized by a decrease in feed intake followed by a decline in growth rates; thus, a fast recovery represents an essential step for proper growth of these animals. Considering that IRMS is a potential tool for non‐destructive sampling and the fact that it provides time‐integrated estimate of assimilated and not just ingested nutrients turned possible its application to evaluate the effects of dietary nucleotides and glutamate on carbon turnover (δ¹³C) in organs of weanling piglets. At day 0, three piglets were slaughtered (prior to diet switch), the remaining eighty‐four piglets weaned at 21‐day‐old were randomly assigned in a complete block design with a 2 × 2 factorial arrangement of treatments (two Nu levels: 0 and 0.1% and two Glu levels: 0 and 1%), being three piglets per treatment slaughtered on trial days 3, 6, 9, 14, 21, 35 and 49. The samples were analysed by IRMS and adjusted to first‐order equation by a non‐linear regression analysis using NLIN of SAS, in order to establish exponential graphics. After that, the turnover data were submitted to analysis of variance using GLM of SAS. The turnover value (t_95%) verified for spleen was faster (p < 0.05) when glutamate was supplemented in diets. For pancreas and liver, the turnover rates were faster (p < 0.05) for the mixture of additives. However, for renal tissue, the turnover rate (t_95%) was greater (p < 0.05) for the free additive diet. The results obtained suggest that the mixture of additives was more efficient to develop the digestive tract at post‐weaning phase, taking into account the functional importance of pancreas and liver for nutrients’ digestion and processing.     

Effect of Maternal Lead Exposure on Craniofacial Ossification in Rat Fetuses and the Role of Antioxidant Therapy

Lead exposure during intrauterine life was found to result in reduced birth weight, impaired skeletal development and post‐natal neurotoxic effects. In this study, the effect of pre‐natal exposure to different doses of lead on the development of craniofacial skeleton in rat fetuses was investigated. Vitamin E was tested as a concomitant treatment, aiming to improve the fetotoxic effects of lead. Positively pregnant female rats were randomly divided into four groups; groups I and II (L250 and L500), exposed to lead acetate in doses of 250 and 500 mg/l respectively, group III (L500 + E), exposed to lead acetate (500 mg/l) wit concomitant vitamin E and group IV (Control) which was given sodium acetate only. All the treatments started from the first day of gestation till the 20th day, where all rats were sacrificed and the fetuses were recovered. Fetuses were processed to alizarin red staining for ossified components. Twenty‐seven bones of the craniofacial skeleton were studied in each fetus where the ossification was scored as being complete, delayed or absent. In all studied fetuses from all groups, changes were found only in eight bones while the remaining craniofacial bones were normally ossified. In affected bones there was a significant decrease in the number of completely ossified bones; associated with a significant increase of both partially ossified and absent bones in L₂₅₀ and L₅₀₀ treated groups when compared to the control group. These differences were more significant in the L₅₀₀ treated group. Giving vitamin E improved the percentage of completely ossified craniofacial bones and decreased the percentage of both partially ossified and absent bones. The most affected bone was presphenoid, then to a lesser extent supraoccipital, squamosal, parietal, interparietal and frontal bone respectively. In conclusion, lead exposure to rats during pregnancy led to varying degrees of fetal growth retardation as well as delayed ossification of some craniofacial bones which were dose dependent and the concomitant supplementation with vitamin E greatly improved the deleterious effect of lead.     

Ultrastructural characteristics of Nematopsis sp. oocysts (Apicomplexa: Porosporidae), a parasite of the clam Meretrix meretrix (Veneridae) from the Arabian Gulf, Saudi Arabia

This paper describes the fine structure of oocysts of Nematopsis sp. (Apicomplexa, Porosporidae) found in the abductor muscles of seawater clams, Meretrix meretrix (Linnaeus, 1758) (Veneridae), collected near the city of Dammam (6 degrees 17'0"N, 50 degrees 12'0"E) in the Arabian Gulf off the coast of Saudi Arabia. Oocysts of an ellipsoidal shape were found among myofibrils of the abductor muscles of infected clams. Each oocyst is composed of an oocyst wall surrounding a single uninucleate vermiform sporozoite located in the lumen of the oocyst wall. The thin oocyst wall (0.70-0.85 microm thick) is composed of homogenous electron-lucent material formed by three layers of equal-thickness. The oocyst wall contains a plano-convex opercular-like structure about 2.5 microm in diameter and 0.75-0.90 microm thick, composed of a homogenous material with moderate electron density. The oocyst is of an ellipsoidal shape and is 15.6 +/- 0.6 microm long and 11.1 +/- 0.7 microm wide. Externally, the oocyst wall is surrounded by a complex dense network of numerous anastomosed microfibrils, which are attached to the oocyst wall, forming 2-3 layers and extending towards the periphery, at some points penetrating amongst the host cells. The myofibrils in some cases show evident aspects of lysis as a consequence of the appearance of lysosome-like vesicles. Lacking knowledge of a complete life cycle and/or molecular data precluded the conclusive identification of this species.