New heterodontosaurid remains from the Canadon Asfalto Formation: cursoriality and the functional importance of the pes in small heterodontosaurids

New ornithischian remains reported here (MPEF-PV 3826) include two complete metatarsi with associated phalanges and caudal vertebrae, from the late Toarcian levels of the Canadon Asfalto Formation. We conclude that these fossil remains represent a bipedal heterodontosaurid but lack diagnostic characters to identify them at the species level, although they probably represent remains of Manidens condorensis, known from the same locality. Histological features suggest a subadult ontogenetic stage for the individual. A cluster analysis based on pedal measurements identifies similarities of this specimen with heterodontosaurid taxa and the inclusion of the new material in a phylogenetic analysis with expanded character sampling on pedal remains confirms the described specimen as a heterodontosaurid. Finally, uncommon features of the digits (length proportions among nonungual phalanges of digit III, and claw features) are also quantitatively compared to several ornithischians, theropods, and birds, suggesting that this may represent a bipedal cursorial heterodontosaurid with gracile and grasping feet and long digits. In particular, the elongated non-terminal pedal phalanges and morphology of digit III resemble features present in arboreal birds, a unique condition found so far among ornithischians

Protein and energy requirements for maintenance and growth in juvenile meagre Argyrosomus regius (Asso,1801) (Sciaenidae)

[EN] The meagre is a fish species of recent interest in aquaculture, because of its fast growth and flesh quality. Nevertheless, it hasn't been studied enough, and feed producers do not have enough information about the nutrient requirements to optimize the feed diets of the meagre. This study measures the growth response of this fish to several amounts of food and gives information about the proportion of protein and energy that should be included in its diet, as well as the recommended amount of food to optimize its growth. The meagre is a carnivorous species and might be a suitable candidate species for the diversification of aquaculture in the Mediterranean region. This is based on its high growth and flesh quality. Nevertheless, there is little information available about its growth rates and nutrient requirements. The objective of this study was to determine the protein and energy requirements of juvenile meagre (Argyrosomus regius). Two trials for different weights of 53 and 188 g were conducted with rations from starvation to apparent satiation with the scope of studying its nutritional needs. In the first trial, the initial mean body weight of the fish was 53 g, and they were fed at feeding rates, measured as a percentage of the body weight, of 0, 0.75, 1.5, 2.5, 3.5, and 4.5%, with two replicates per treatment. In a second trial, another group with approximately 188 g of initial body weight was fed at feeding rates of 0, 0.5, 1.5, and 2.5%, with two replicates per treatment. The optimum thermal growth coefficient was obtained with a feed intake of 2.2% day(-1) in trial A and 1.73% day(-1) in trial B. The digestible protein (DP) intake for maintenance was determined as 0.57 g kg(-0.7) day(-1), the DP intake for maximum growth was 6.0 g kg(-0.7) day(-1), and the point for maximum efficiency in protein retention was 1.8 g kg(-0.7) day(-1). The requirement for digestible energy (DE) intake for maintenance was recorded at 25.4 kJ kg(-0.82) day(-1), the DE intake to maximize growth was 365 kJ kg(-0.82) day(-1), and the point for maximum efficiency in energy retention occurs with a digestible energy intake of 93 kJ kg(-0.82) day(-1). The requirements and retention efficiency of protein and energy in Argyrosomus regius tend to be within the range other fish species. The maintenance needs are in agreement with species with low voluntary activity and growth requirements in agreement with fast-growth species.This research was funded by grants from the Planes Nacionales de Acuicultura (JACUMAR) in Spain.Jauralde García, I.; Velazco-Vargas, J.; Tomas-Vidal, A.; Jover Cerda, M.; Martínez-Llorens, S. (2021). Protein and energy requirements for maintenance and growth in juvenile meagre Argyrosomus regius (Asso,1801) (Sciaenidae). Animals. 11(1):1-15. https://doi.org/10.3390/ani11010077S115111Chatzifotis, S., Panagiotidou, M., Papaioannou, N., Pavlidis, M., Nengas, I., & Mylonas, C. C. (2010). Effect of dietary lipid levels on growth, feed utilization, body composition and serum metabolites of meagre (Argyrosomus regius) juveniles. Aquaculture, 307(1-2), 65-70. doi:10.1016/j.aquaculture.2010.07.002EL-Shebly, A. A., El-Kady, M. A. H., Hussin, A. B., & Hossain, M. Y. (2007). Preliminary Observations on the Pond Culture of Meagre, Argyrosomus regius (Asso, 1801) (Sciaenidae) in Egypt. Journal of Fisheries and Aquatic Science, 2(5), 345-352. doi:10.3923/jfas.2007.345.352ESTÉVEZ, A., TREVIÑO, L., KOTZAMANIS, Y., KARACOSTAS, I., TORT, L., & GISBERT, E. (2010). Effects of different levels of plant proteins on the ongrowing of meagre (Argyrosomus regius) juveniles at low temperatures. Aquaculture Nutrition, 17(2), e572-e582. doi:10.1111/j.1365-2095.2010.00798.xPoli, B. M., Parisi, G., Zampacavallo, G., Iurzan, F., Mecatti, M., Lupi, P., & Bonelli, A. (2003). Aquaculture International, 11(3), 301-311. doi:10.1023/a:1024840804303Roo, J., Hernández-Cruz, C. M., Borrero, C., Schuchardt, D., & Fernández-Palacios, H. (2010). Effect of larval density and feeding sequence on meagre (Argyrosomus regius; Asso, 1801) larval rearing. Aquaculture, 302(1-2), 82-88. doi:10.1016/j.aquaculture.2010.02.015Chatzifotis, S., Panagiotidou, M., & Divanach, P. (2011). Effect of protein and lipid dietary levels on the growth of juvenile meagre (Argyrosomus regius). Aquaculture International, 20(1), 91-98. doi:10.1007/s10499-011-9443-yAlvarez-González, C. ., Civera-Cerecedo, R., Ortiz-Galindo, J. ., Dumas, S., Moreno-Legorreta, M., & Grayeb-Del Alamo, T. (2001). Effect of dietary protein level on growth and body composition of juvenile spotted sand bass, Paralabrax maculatofasciatus, fed practical diets. Aquaculture, 194(1-2), 151-159. doi:10.1016/s0044-8486(00)00512-3Chong, A. S. ., Ishak, S. D., Osman, Z., & Hashim, R. (2004). Effect of dietary protein level on the reproductive performance of female swordtails Xiphophorus helleri (Poeciliidae). Aquaculture, 234(1-4), 381-392. doi:10.1016/j.aquaculture.2003.12.003El-Sayed, A.-F. M., & Kawanna, M. (2008). Effects of dietary protein and energy levels on spawning performance of Nile tilapia (Oreochromis niloticus) broodstock in a recycling system. Aquaculture, 280(1-4), 179-184. doi:10.1016/j.aquaculture.2008.04.030Lee, S.-M., Jeon, I. G., & Lee, J. Y. (2002). Effects of digestible protein and lipid levels in practical diets on growth, protein utilization and body composition of juvenile rockfish (Sebastes schlegeli). Aquaculture, 211(1-4), 227-239. doi:10.1016/s0044-8486(01)00880-8Zhang, J., Zhou, F., Wang, L., Shao, Q., Xu, Z., & Xu, J. (2010). Dietary Protein Requirement of Juvenile Black Sea Bream, Sparus macrocephalus. Journal of the World Aquaculture Society, 41, 151-164. doi:10.1111/j.1749-7345.2010.00356.xTibbetts, S. M., Lall, S. P., & Anderson, D. M. (2000). Dietary protein requirement of juvenile American eel (Anguilla rostrata) fed practical diets. Aquaculture, 186(1-2), 145-155. doi:10.1016/s0044-8486(99)00363-4Kaushik, S. J., & Seiliez, I. (2010). Protein and amino acid nutrition and metabolism in fish: current knowledge and future needs. Aquaculture Research, 41(3), 322-332. doi:10.1111/j.1365-2109.2009.02174.xGunasekera, R. M., De Silva, S. S., Collins, R. A., Gooley, G., & Ingram, B. A. (2000). Effect of dietary protein level on growth and food utilization in juvenile Murray codMaccullochella peelii peelii(Mitchell). Aquaculture Research, 31(2), 181-187. doi:10.1046/j.1365-2109.2000.00417.xBooth, M. A., Allan, G. L., & Pirozzi, I. (2010). Estimation of digestible protein and energy requirements of yellowtail kingfish Seriola lalandi using a factorial approach. Aquaculture, 307(3-4), 247-259. doi:10.1016/j.aquaculture.2010.07.019Jauralde, I., Martínez-Llorens, S., Tomás, A., & Jover, M. (2016). Protein deposition and energy recovery in gilthead sea bream (Sparus aurata): Evaluation of nutritional requirements. Aquaculture, 464, 65-73. doi:10.1016/j.aquaculture.2016.06.006Lupatsch, I., Kissil, G. W., Sklan, D., & Pfeffer, E. (1998). Energy and protein requirements for maintenance and growth in gilthead seabream (Sparus aurata L.). Aquaculture Nutrition, 4(3), 165-173. doi:10.1046/j.1365-2095.1998.00065.xLupatsch, Kissil, Sklan, & Pfeffer. (2001). Effects of varying dietary protein and energy supply on growth, body composition and protein utilization in gilthead seabream (Sparus aurataL.). Aquaculture Nutrition, 7(2), 71-80. doi:10.1046/j.1365-2095.2001.00150.xPeres, H., & Oliva-Teles, A. (2005). Protein and Energy Metabolism of European Seabass (Dicentrarchus labrax) Juveniles and Estimation of Maintenance Requirements. Fish Physiology and Biochemistry, 31(1), 23-31. doi:10.1007/s10695-005-4586-2Lupatsch, I., & Kissil, G. W. (2005). Feed formulations based on energy and protein demands in white grouper (Epinephelus aeneus). Aquaculture, 248(1-4), 83-95. doi:10.1016/j.aquaculture.2005.03.004Pirozzi, I., Booth, M. A., & Allan, G. L. (2008). Protein and energy utilization and the requirements for maintenance in juvenile mulloway (Argyrosomus japonicus). Fish Physiology and Biochemistry, 36(1), 109-121. doi:10.1007/s10695-008-9296-0McGoogan, B. B., & Gatlin, D. M. (1998). Metabolic Requirements of Red Drum, Sciaenops ocellatus, for Protein and Energy Based on Weight Gain and Body Composition. The Journal of Nutrition, 128(1), 123-129. doi:10.1093/jn/128.1.123GLENCROSS, B. D. (2009). Reduced water oxygen levels affect maximal feed intake, but not protein or energy utilization efficiency of rainbow trout (Oncorhynchus mykiss). Aquaculture Nutrition, 15(1), 1-8. doi:10.1111/j.1365-2095.2007.00562.xGlencross, B., Hawkins, W., Evans, D., Rutherford, N., Dods, K., McCafferty, P., & Sipsas, S. (2007). Evaluation of the influence of drying process on the nutritional value of lupin protein concentrates when fed to rainbow trout (Oncorhynchus mykiss). Aquaculture, 265(1-4), 218-229. doi:10.1016/j.aquaculture.2007.01.040Rodehutscord, M., & Pfeffer, E. (1999). Maintenance requirement for digestible energy and efficiency of utilisation of digestible energy for retention in rainbow trout, Oncorhynchus mykiss. Aquaculture, 179(1-4), 95-107. doi:10.1016/s0044-8486(99)00155-6Booth, M. A., & Allan, G. L. (2003). Utilization of digestible nitrogen and energy from four agricultural ingredients by juvenile silver perch Bidyanus bidyanus. Aquaculture Nutrition, 9(5), 317-326. doi:10.1046/j.1365-2095.2003.00259.xHatlen, B., Helland, S. J., & Grisdale-Helland, B. (2007). Energy and nitrogen partitioning in 250 g Atlantic cod (Gadus morhua L.) given graded levels of feed with different protein and lipid content. Aquaculture, 270(1-4), 167-177. doi:10.1016/j.aquaculture.2007.04.001GLENCROSS, B. D. (2008). A factorial growth and feed utilization model for barramundi,Lates calcariferbased on Australian production conditions. Aquaculture Nutrition, 14(4), 360-373. doi:10.1111/j.1365-2095.2007.00543.xHelland, S. J., Hatlen, B., & Grisdale-Helland, B. (2010). Energy, protein and amino acid requirements for maintenance and efficiency of utilization for growth of Atlantic salmon post-smolts determined using increasing ration levels. Aquaculture, 305(1-4), 150-158. doi:10.1016/j.aquaculture.2010.04.013Fournier, V., Gouillou-Coustans, M. F., Métailler, R., Vachot, C., Guedes, M. J., Tulli, F., … Kaushik, S. J. (2002). Protein and arginine requirements for maintenance and nitrogen gain in four teleosts. British Journal of Nutrition, 87(5), 459-469. doi:10.1079/bjn2002564Bureau, D. P., Hua, K., & Cho, C. Y. (2006). Effect of feeding level on growth and nutrient deposition in rainbow trout (Oncorhynchus mykiss Walbaum) growing from 150 to 600 g. Aquaculture Research, 37(11), 1090-1098. doi:10.1111/j.1365-2109.2006.01532.xAtkinson, J. L., Hilton, J. W., & Slinger, S. J. (1984). Evaluation of Acid-Insoluble Ash as an Indicator of Feed Digestibility in Rainbow Trout (Salmo gairdneri). Canadian Journal of Fisheries and Aquatic Sciences, 41(9), 1384-1386. doi:10.1139/f84-170Watanabe, K., Ura, K., Yada, T., Kiron, V., Satoh, S., & Watanabe, T. (2000). Energy and protein requirements of yellowtail for maximum growth and maintenance of body weight. Fisheries Science, 66(6), 1053-1061. doi:10.1046/j.1444-2906.2000.00168.xDumas, A., France, J., & Bureau, D. P. (2007). Evidence of three growth stanzas in rainbow trout (Oncorhynchus mykiss) across life stages and adaptation of the thermal-unit growth coefficient. Aquaculture, 267(1-4), 139-146. doi:10.1016/j.aquaculture.2007.01.041Jauralde, I., Martínez-Llorens, S., Tomás, A., Ballestrazzi, R., & Jover, M. (2011). A proposal for modelling the thermal-unit growth coefficient and feed conversion ratio as functions of feeding rate for gilthead sea bream (Sparus aurata,L.) in summer conditions. Aquaculture Research, 44(2), 242-253. doi:10.1111/j.1365-2109.2011.03027.xMayer, P., Estruch, V. D., & Jover, M. (2012). A two-stage growth model for gilthead sea bream (Sparus aurata) based on the thermal growth coefficient. Aquaculture, 358-359, 6-13. doi:10.1016/j.aquaculture.2012.06.016Panettieri, V., Chatzifotis, S., Messina, C. M., Olivotto, I., Manuguerra, S., Randazzo, B., … Piccolo, G. (2020). Honey Bee Pollen in Meagre (Argyrosomus regius) Juvenile Diets: Effects on Growth, Diet Digestibility, Intestinal Traits, and Biochemical Markers Related to Health and Stress. Animals, 10(2), 231. doi:10.3390/ani10020231Knibb, W. (2000). Genetic improvement of marine fish - which method for industry? Aquaculture Research, 31(1), 11-23. doi:10.1046/j.1365-2109.2000.00393.xWatanabe, K., Hara, Y., Ura, K., Yada, T., Kiron, V., Satoh, S., & Watanabe, T. (2000). Energy and protein requirements for maximum growth and maintenance of bodyweight of yellowtail. Fisheries Science, 66(5), 884-893. doi:10.1046/j.1444-2906.2000.00143.xLupatsch, I., Kissil, G. W., & Sklan, D. (2001). Optimization of feeding regimes for European sea bass Dicentrarchus labrax: a factorial approach. Aquaculture, 202(3-4), 289-302. doi:10.1016/s0044-8486(01)00779-7Arshad Hossain, M., Almatar, S. M., & James, C. M. (2010). Optimum Dietary Protein Level for Juvenile Silver Pomfret, Pampus argenteus (Euphrasen). Journal of the World Aquaculture Society, 41(5), 710-720. doi:10.1111/j.1749-7345.2010.00413.xSandberg, F. B., Emmans, G. C., & Kyriazakis, I. (2005). Partitioning of limiting protein and energy in the growing pig: testing quantitative rules against experimental data. British Journal of Nutrition, 93(2), 213-224. doi:10.1079/bjn20041322Sánchez-Lozano, N. B., Martínez-Llorens, S., Tomás-Vidal, A., & Cerdá, M. J. (2009). Effect of high-level fish meal replacement by pea and rice concentrate protein on growth, nutrient utilization and fillet quality in gilthead seabream (Sparus aurata, L.). Aquaculture, 298(1-2), 83-89. doi:10.1016/j.aquaculture.2009.09.028SÁNCHEZ-LOZANO, N. B., MARTÍNEZ-LLORENS, S., TOMÁS-VIDAL, A., & JOVER CERDÁ, M. (2010). Amino acid retention of gilthead sea bream (Sparus aurata, L.) fed with pea protein concentrate. Aquaculture Nutrition, 17(2), e604-e614. doi:10.1111/j.1365-2095.2010.00803.xHillestad, M., & Johnsen, F. (1994). High-energy/low-protein diets for Atlantic salmon: effects on growth, nutrient retention and slaughter quality. Aquaculture, 124(1-4), 109-116. doi:10.1016/0044-8486(94)90366-2Shearer, K. D. (1994). Factors affecting the proximate composition of cultured fishes with emphasis on salmonids. Aquaculture, 119(1), 63-88. doi:10.1016/0044-8486(94)90444-

Effects of Eco-Organic Feed on Growth Performance, Biometric Indices, and Nutrient Retention of Gilthead Seabream (Sparus aurata)

[EN] This study examined how eco-organic feed affects the growth performance, nutrient efficiency, feed utilisation, and body composition of gilthead seabream. Six different diets were tested, including a control diet (CONT) without organic ingredients and four diets with 100% organic ingredients: trout (TRO), seabass (SBS), poultry (POU), and mix (MIX), along with a control organic diet (ORG) containing organic ingredients and 30% fishmeal. The experiment lasted 70 days, and the fish were fed twice a day, starting with an initial weight of 60.5 g. The results showed that the highest growth rates were observed in fish fed the ORG and CONT diets containing fishmeal. Conversely, the POU diet resulted in the lowest growth rate, survival rate, and highest value for feed conversion ratio (FCR). Almost all essential amino acid efficiency values were high in fish fed the ORG and CONT diets. Still, significant differences were noted in the retention efficiency of fatty acids across all diets. The retention efficiency was higher in the CONT diet, followed by the ORG diet. However, the economic conversion rate was lower for CONT, SBS, TRO, and MIX. Overall, using organic diets of animal origin impacted the growth performance of gilthead seabream, but it is still a promising approach.This project had been developed with the collaboration of the Biodiversity Foundation (Spanish Ministry for Ecological Transition and the Demographic Challenge), through the Pleamar Program, co-financed by the European Maritime and Fisheries Fund (EMFF). A full scholarship from the Ministry of Higher Education of the Arab Republic of Egypt funds the researcher Eslam TefalTefal, E.; Tomas-Vidal, A.; Martínez-Llorens, S.; Jauralde García, I.; Peñaranda, D.; Jover Cerda, M. (2023). Effects of Eco-Organic Feed on Growth Performance, Biometric Indices, and Nutrient Retention of Gilthead Seabream (Sparus aurata). Sustainability. 15(14):1-16. https://doi.org/10.3390/su151410750116151

Protocolo para la preparación de secciones delgadas sin descalcificar de tejido óseo humano

La obtención de cortes delgados de restos óseos correspondiente tanto a materiales fósiles como arqueológicos suele ser una tarea poco sencilla, lo cual conlleva generalmente un elevado costo y tiempos prolongados. Por otra parte, la formación de un laboratorio con las características necesarias para la realización de protocolos ya establecidos es sumamente costosa. En este trabajo se presenta un protocolo alternativo para la obtención de cortes delgados de muestras óseas humanas de sitios arqueológicos con el objetivo de aplicarlo al análisis microestructural. Los procedimientos de obtención de cortes delgados que aquí se detallan están al alcance de laboratorios de reciente formación. En primer lugar, se midió y se registró con imágenes digitales un fémur, una tibia y una primera costilla de un individuo adulto procedente de un cementerio contemporáneo. Luego se extrajeron fragmentos de 1 cm de hueso. Con el material se confeccionaron réplicas y por último se obtuvieron los cortes delgados. Los resultados obtenidos fueron óptimos para la observación de las características microestructurales del hueso. Con los ajustes adecuados (tiempo de fraguado, granulometría del abrasivo de acuerdo a la dureza del material, impregnación, etc.) este protocolo podría ser utilizado para diversos tipos de muestras actuales, arqueológicas e incluso fósiles

Influence of diet and feeding strategy on the performance of nitrifying trickling filter, oxygen consumption and ammonia excretion of gilthead sea bream (Sparus aurata) raised in recirculating aquaculture systems

[EN] Gilthead sea bream (Sparus aurata) was raised in six individual recirculating aquaculture systems (RAS) whose bioflters¿ performance was analyzed. Fish were fed with three diferent diets (a control diet, a fshmeal-based diet (FM), and a plant meal-based diet (VM)) and with three diferent feeding strategies (manual feeding to apparent satiation, automatic feeding with restricted ration, and auto-demand feeding). For every combination of diet and feeding strategy, the mean oxygen consumption, ammonia excretion, and ammonia removal rate were determined. Fish fed with the VM diet consumed the most oxygen (20.06±1.80 gO2 consumed kg¿1 day¿1). There were signifcant diferences in ammonia excretion depending on the protein content and protein efciency of the diet, as well as depending on feeding strategy, which in turn afected ammonia removal rates. Fish fed by auto-demand feeders led to the highest mean ammonia removal rate (0.10 gN-TAN removed m¿2 biofltration area day¿1), while not leading to peaks of high ammonia concentration in water, which preserve fsh welfare and growth.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Research is funded by the national project ¿Design of a recirculating aquaculture system for aquaculture plants (2011¿2014),¿ by the Ministry of Science and Innovation, Spain, as well as by a grant financed by Generalitat Valenciana, IDIFEDER/2020/029, and by the project ¿Recirculating aquaculture systems¿ by Universitat Politècnica de València. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Godoy-Olmos, S.; Jauralde García, I.; Monge-Ortiz, R.; Milián Sorribes, MC.; Jover Cerda, M.; Tomas-Vidal, A.; Martínez-Llorens, S. (2022). Influence of diet and feeding strategy on the performance of nitrifying trickling filter, oxygen consumption and ammonia excretion of gilthead sea bream (Sparus aurata) raised in recirculating aquaculture systems. Aquaculture International. 30(2):581-606. https://doi.org/10.1007/s10499-021-00821-358160630

Gregariousness in the giant sloth Lestodon (Xenarthra): multi-proxy approach of a bonebed from the Last Maximum Glacial of Argentine Pampas

Megamammals constituted an important component in the Pleistocene faunal communities of South America. Paleobiological and paleoecological studies involving different megamammal taxa have increased significantly in the last years, but there are still several poorly-known issues of its life history. In this work, we analyze an assemblage composed of 13 individuals of different ontogenetic stages, and possibly different sex, belonging to the giant ground sloth Lestodon armatus (Xenarthra, Folivora), recovered from Playa del Barco site (Pampean Region, Argentina). A dating of 19,849 years Cal BP allows assigning this assemblage to a period of the MIS (Marine Isotope Stage) 2 related to the end of the Last Glacial Maximum. Based on multiple lines of research (e.g. taphonomy, paleopathology, osteohistology, isotopy), we interpret the origin of the assemblage and diverse paleobiological and paleoecological aspects (e.g. social behavior, ontogenetic changes, sexual dimorphism, diseases, resource and habitat use, trophic relationships) of L. armatus. Evidence suggests that the assemblage was formed by a local single event of catastrophic mortality, which affected different members of a social group. This record represents the first accurate evidence of gregariousness for this ground sloth, providing new data on a poorly-known behavior among extinct Folivora

Brucella-Salmonella lipopolysaccharide chimeras are less permeable to hydrophobic probes and more sensitive to cationic peptides and EDTA than are their native Brucella sp. counterparts

A rough (R) Brucella abortus 45/20 mutant was more sensitive to the bactericidal activity of polymyxin B and lactoferricin B than was its smooth (S) counterpart but considerably more resistant than Salmonella montevideo. The outer membrane (OM) and isolated lipopolysaccharide (LPS) of S. montevideo showed a higher affinity for these cationic peptides than did the corresponding B. abortus OM and LPS. We took advantage of the moderate sensitivity of R B. abortus to cationic peptides to construct live R B. abortus-S-LPS chimeras to test the activities of polymyxin B, lactoferricin B, and EDTA. Homogeneous and abundant peripheral distribution of the heterologous S-LPS was observed on the surface of the chimeras, and this coating had no effect on the viability or morphology of the cells. When the heterologous LPS corresponded to the less sensitive bacterium S B. abortus S19, the chimeras were more resistant to cationic peptides; in contrast, when the S-LPS was from the more sensitive bacterium S. montevideo, the chimeras were more susceptible to the action of peptides and EDTA. A direct correlation between the amount of heterologous S-LPS on the surface of chimeric Brucella cells and peptide sensitivity was observed. Whereas the damage produced by polymyxin B in S. montevideo and B. abortus-S. montevideo S-LPS chimeras was manifested mainly as OM blebbing and inner membrane rolling, lactoferricin B caused inner membrane detachment, vacuolization, and the formation of internal electron-dense granules in these cells. Native S and R B. abortus strains were permeable to the hydrophobic probe N-phenyl-1-naphthylamine (NPN). In contrast, only reduced amounts of NPN partitioned into the OMs of the S. montevideo and B. abortus-S. montevideo S-LPS chimeras. Following peptide exposure, accelerated NPN uptake similar to that observed for S. montevideo was detected for the B. abortus-S. montevideo LPS chimeras. The partition of NPN into native or EDTA-, polymyxin B-, or lactoferricin B-treated LPS micelles of S. montevideo or B. abortus mimicked the effects observed with intact cells, and this was confirmed by using micelle hybrids of B. abortus and S. montevideo LPSs. The results showed that LPS is the main cause of B. abortus' resistance to bactericidal cationic peptides, the OM-disturbing action of divalent cationic chelants, and OM permeability to hydrophobic substances. It is proposed that these three features are related to the ability of Brucella bacteria to multiply within phagocytes

The outer membrane of Brucella ovis shows increased permeability to hydrophobic probes and is more susceptible to cationic peptides than are the outer membranes of mutant rough Brucella abortus strains

The permeability of the outer membrane (OM) to hydrophobic probes and its susceptibility to bactericidal cationic peptides were investigated for natural rough Brucella ovis and for mutant rough Brucella abortus strains. The OM of B. ovis displayed an abrupt and faster kinetic profile than rough B. abortus during the uptake of the hydrophobic probe N-phenyl-naphthylamine. B. ovis was more sensitive than rough B. abortus to the action of cationic peptides. Bactenecins 5 and 7 induced morphological alterations on the OMs of both rough Brucella strains. B. ovis lipopolysaccharide (LPS) captured considerably more polymyxin B than LPSs from both rough and smooth B. abortus strains. Polymyxin B, poly-L-lysine, and poly-L-ornithine produced a thick coating on the surfaces of both strains, which was more evident in B. ovis than in rough B. abortus. The distinct functional properties of the OMs of these two rough strains correlate with some structural differences of their OMs and with their different biological behaviors in animals and culture cells

123I-MIBG cardiac uptake and smell identification in parkinsonian patients with LRRK2 mutations

Reduced uptake of 123I- metaiodobenzylguanidine (MIBG) on cardiac gammagraphy and impaired odor identification are markers of neurodegenerative diseases with Lewy bodies (LB) as a pathological hallmark, such as idiopathic Parkinson’s disease (IPD). LRRK2 patients present with a clinical syndrome indistinguishable from IPD, but LB have not been found in some cases. Patients with such mutations could behave differently than patients with IPD with respect to MIBG cardiac uptake and olfaction. We studied 14 LRRK2 patients, 14 IPD patients matched by age, gender, disease duration and severity, and 13 age and gender matched control subjects. Olfaction was analyzed through the University of Pennsylvania Smell Identification Test (UPSIT). MIBG cardiac uptake was evaluated through the H/M ratio. The late H/M was 1.44 ± 0.31 for LRRK2 patients, 1.19 ± 0.15 for PD patients, and 1.67 ± 0.16 for control subjects. LRRK2 patients presented lower but not statistically significant MIBG cardiac uptake than controls (p = 0.08) and significant higher uptake than PD patients (p = 0.04). UPSIT mean scores were 21.5 ± 7.3 for LRRK2 patients, 18.7 ± 6.2 for IPD patients and 29.7 ± 5.7 for control subjects. UPSIT score was lower in both LRRK2 and PD than in controls. In LRRK2 patients a positive correlation was found between myocardial MIBG uptake and UPSIT scores, (R = 0.801, p < 0.001). In LRRK2 patients, MIBG cardiac uptake was less impaired than in PD; a positive correlation between MIBG cardiac uptake and UPSIT scores was observed. As MIBG cardiac reduced uptake and impaired odor identification are markers of LB pathology, this findings may represent neuropathological heterogeneity among LRRK2 patients

Evaluation of the effects of erythritol on gene expression in Brucella abortus

Bacteria of the genus Brucella have the unusual capability to catabolize erythritol and this property has been associated with their virulence mainly because of the presence of erythritol in bovine foetal tissues and because the attenuated S19 vaccine strain is the only Brucella strain unable to oxydize erythritol. In this work we have analyzed the transcriptional changes produced in Brucella by erythritol by means of two high throughput approaches: RNA hybridization against a microarray containing most of Brucella ORF's constructed from the Brucella ORFeome and next generation sequencing of Brucella mRNA in an Illumina GAIIx platform. The results obtained showed the overexpression of a group of genes, many of them in a single cluster around the ery operon, able to co-ordinately mediate the transport and degradation of erythritol into three carbon atoms intermediates that will be then converted into fructose-6P (F6P) by gluconeogenesis. Other induced genes participating in the nonoxidative branch of the pentose phosphate shunt and the TCA may collaborate with the ery genes to conform an efficient degradation of sugars by this route. On the other hand, several routes of amino acid and nucleotide biosynthesis are up-regulated whilst amino acid transport and catabolism genes are down-regulated. These results corroborate previous descriptions indicating that in the presence of erythritol, this sugar was used preferentially over other compounds and provides a neat explanation of the the reported stimulation of growth induced by erythritol