A theropod trackway providing evidence of a pathological foot from the exceptional locality of Las Hoyas (upper Barremian, Serranía de Cuenca, Spain)

We describe a trackway (LH-Mg-10-16) occurring in laminated carbonated limestones of the Las Hoyas locality, Serranía de Cuenca, Spain. It is unmistakably a large theropod dinosaur trackway encompassing two unusual aspects, namely, wide-steps, and a set of equally deformed left footprints (with a dislocated digit). The layer also preserves other vertebrate trails (fish Undichna) and different impressions in the sediment. To address these complex settings, we devised a multidisciplinary approach, including the ichnological and taphonomical descriptions, characterisation of the rock lithofacies using thin-sections, 3D structured-light digitalisation with a high precision of 200-400 μm, and a geometric morphometric comparison with a large sample of bipedal dinosaur trackways. Sedimentary analyses showed that the trackway was produced in a humid, benthonic microbial mat, the consistency and plasticity of which enabled the preservation of the details of the movement of the animal. The results of the geometric analysis indicate that the 'wide-steps' of the trackway is not unusual compared to other trackways, providing evidence that it was made by a single individual with an estimated hip height approximately 2 m. Analogous pathologies in extant archosaurs that yield the combination of wide steps and deformed digits in the same trackway were considered. All results mutually support the hypothesis that a large theropod dinosaur, with a pathological foot, generated the trackway as it crossed an area of shallow water while slowly walking towards the main water source, thus stepping steadily over the benthonic mat over which multiple fish were swimming

Delineating ecologically significant taxonomic units from global patterns of marine picocyanobacteria

Prochlorococcus and Synechococcus are the two most abundant and widespread phytoplankton in the global ocean. To better understand the factors controlling their biogeography, a reference database of the high-resolution taxonomic marker petB, encoding cytochrome b6, was used to recruit reads out of 109 metagenomes from the Tara Oceans expedition. An unsuspected novel genetic diversity was unveiled within both genera, even for the most abundant and well-characterized clades, and 136 divergent petB sequences were successfully assembled from metagenomic reads, significantly enriching the reference database. We then defined Ecologically Significant Taxonomic Units (ESTUs)—that is, organisms belonging to the same clade and occupying a common oceanic niche. Three major ESTU assemblages were identified along the cruise transect for Prochlorococcus and eight for Synechococcus. Although Prochlorococcus HLIIIA and HLIVA ESTUs codominated in irondepleted areas of the Pacific Ocean, CRD1 and the yet-to-be cultured EnvB were the prevalent Synechococcus clades in this area, with three different CRD1 and EnvB ESTUs occupying distinct ecological niches with regard to iron availability and temperature. Sharp community shifts were also observed over short geographic distances—for example, around the Marquesas Islands or between southern Indian and Atlantic Oceans—pointing to a tight correlation between ESTU assemblages and specific physico-chemical parameters. Together, this study demonstrates that there is a previously overlooked, ecologically meaningful, fine-scale diversity within some currently defined picocyanobacterial ecotypes, bringing novel insights into the ecology, diversity, and biology of the two most abundant phototrophs on Earth

Microbiological, pathological and microelement analyses in vicuñas affected with "dandruff"

Se describen 75 estudios histopatológicos en biopsias de piel (33 afectados y 42 no afectados con “caspa”), 85 análisis microbiológicos en raspados de piel (44 afectados y 41 no afectados) y 70 determinaciones séricas de zinc, selenio, cobre y molibdeno (41 afectados y 29 no afectados) de tres poblaciones de vicuñas silvestres capturadas en “Chakus” en el 2009 en las comunidades campesinas de Huaytará, Ayaví, Santa Rosa de Tambo y en una población captiva multicomunal, en Huancavelica. Los animales afectados no tenían alteraciones clínicas, pero los vellones a la postesquila presentaron escamas blanquecinas dispersas o acumuladas y fuertemente adheridas, usualmente, al dorso lateral y algunas veces por todo el vellón. Todas las muestras de piel, con mayor severidad en las afectadas, mostraron moderada hiperqueratosis ortoqueratótica laminar asociada con dermatosis inespecífica, moderada-severa dilatación de folículos pilosos y moderada-severa atrofia de vaina interna de la raíz folicular pero con ausencia de agentes patógenos e inflamación. El 63.3% (28/44) de raspados de pieles afectadas y el 41.5% (17/ 41) de las no afectadas contenían especies saprofíticas de Ulocladium spp., Penicillum spp., Hialofomicetos, Geotrichum candidum y Aspergilus flavus. Los niveles sanguíneos, en las 70 muestras (afectados y no afectados) presentaron 10 veces la concentración esperada para selenio, principalmente en vicuñas captivas en el área multicomunal (afectados 3.23 ± 1.31 μg/mL y no afectados 3.56 ± 2.27 μg/mL), posiblemente debido al sobrepastoreo de los pastizales con presencia de especies seleníferas de Astragalus spp. (“garbanzo” o “garbancillo”). Todos los animales mostraron deficiencia de cobre y los animales afectados de Huaytará y todos los de Santa Rosa de Tambo presentaron deficiencia de zinc.In recent years important economic losses have resulted from what is described as “dandruff” in vicuña fiber. With the goal of analyzing the possible cause/s was conducted an histopathological analysis of 75 skin biopsies (33 affected/42 unaffected), microbiological analysis of 85 skin/fiber scrapings (44 affected/41 unaffected), and microelement analysis (zinc, selenium, copper, molybdenum) of 70 serum samples (41 affected/ 29 unaffected), collected from three wild populations in the communities of Huaytará, Ayaví and Santa Rosa de Tambo, Huancavelica, Peru, as well as from the captive herd held jointly by these communities. The affected vicuñas were clinically normal and the presence of “dandruff” was generally detected after shearing. In these fleeces, white scales scattered or accumulated and firmly adhered to the fibers were found, especially on the flanks and backs of the animals, but also widely dispersed throughout the fleece. Histopathological analysis of the skin biopsies revealed that both affected and unaffected animals had moderate to severe dermatosis (hyperkeratosis – orthokeratosis), with moderate to severe atrophy of the inner root sheath of the follicle, but without evidence of inflammation. Microbiological analysis determined the presence of fungus species in 63.3% (28/44) of the affected and 41.5% (17/41) of unaffected animals, including Ulocladium spp., Penicillum spp., Hialofomicetos, Geotrichum candidum and Aspergilus flavus. Microelement analysis revealed 10 fold selenium concentration as compared to normal values, especially in the captive population (affected: 3.23 ± 1.31 μg/ mL; unaffected: 3.56 ± 2.27 μg/mL) possibly due to overgrazing of pastures with presence of Astragalus spp., a common seleniferous plant in the region. All animals showed cooper deficiency. Also, all animals from Santa Rosa de Tambo and affected animals from Huaytará were zinc deficient

CONTRIBUTION TO THE STUDY OF GASTROINTESTINAL PARASITISM IN THE GUANACO (LAMA GUANICOE CACSILENSIS)

El propósito del presente estudio fue identificar las especies de parásitosgastrointestinales que afectan al guanaco peruano y determinar los niveles de parasitismode las poblaciones evaluadas. Se obtuvieron 132 muestras de heces frescas deguanacos silvestres pertenecientes a nueve poblaciones ubicadas en seis departamentosdel Perú: Comunidad Campesina de Huallhua (Ayacucho), Reserva Nacional de Calipuy(La Libertad), Comunidad Campesina de Chavín (Ica), Reserva Nacional Salinas y AguadaBlanca y distritos de Machaguay y Yarabamba (Arequipa), distrito de Quilahuani yComunidad Campesina de Vila Vilani (Tacna), y distrito de La Capilla (Moquegua). Lasmuestras fueron procesadas mediante técnicas coproparasitológicas de flotación, sedimentación,cultivo de larvas, Baerman y biometría de larvas y ooquistes. Se identificaronocho especies de nematodos: Graphinema aucheniae, Bunostomun sp., Ostertagia sp.,Trichuris sp, Cooperia sp., Nematodirus sp., Mazamastrongylus peruvianus yTrichostrongylus sp. y cuatro especies de Eimeria: E. lamae, E. alpacae, E. punoensis yE. macusaniensis. Todas las poblaciones se encontraban con al menos un guanacoparasitado, presentando en general cargas bajas y variando las frecuencias de parasitismogastrointestinal de una población a otra, dependiendo del hábitat y de la proximidada herbívoros domésticos.The aim of this study was to identify the species of gastrointestinal parasites affecting the Peruvian guanaco and to determine the levels of parasitism in the populations under evaluation. For this purpose, 132 fresh faecal samples were collected from nine populations of wild guanacos located in six departments of Peru: Huallhua Community in Ayacucho; Calipuy National Reserve in La Libertad; Chavín community in Ica; Salinas and Aguada Blanca National Reserve, and Machaguay and Yarabamba districts in Arequipa, Quilahuani district and Vila Vilani community in Tacna, and La Capilla district in Moquegua. Samples were processed by the coproparasitological techniques of flotation, sedimentation, larvae culture, and Baerman, and biometry of larvae and oocysts. Eight species of nematodes were identified: Graphinema aucheniae, Bunostomun spp., Ostertagia spp., Trichuris spp., Cooperia spp., Nematodirus spp., Mazamastrongylus peruvianus and Trichostrongylus spp., and four Eimeria species: E. lamae, E. alpacae, E. punoensis and E. macusaniensis. All guanaco populations had at least one animal with parasites, showing low parasite burden in general, and with a variation in the frequency of gastrointestinal parasitism from one population to another, depending on the habitat and the proximity to other domestic herbivores

Variations of training load, monotony, and strain and dose-response relationships with maximal aerobic speed, maximal oxygen uptake, and isokinetic strength in professional soccer players

This study aimed to identify variations in weekly training load, training monotony, and training strain across a 10-week period (during both, pre- and in-season phases); and to analyze the dose-response relationships between training markers and maximal aerobic speed (MAS), maximal oxygen uptake, and isokinetic strength. Twenty-seven professional soccer players (24.9±3.5 years old) were monitored across the 10-week period using global positioning system units. Players were also tested for maximal aerobic speed, maximal oxygen uptake, and isokinetic strength before and after 10 weeks of training. Large positive correlations were found between sum of training load and extension peak torque in the right lower limb (r = 0.57, 90%CI[0.15;0.82]) and the ratio agonist/antagonist in the right lower limb (r = 0.51, [0.06;0.78]). It was observed that loading measures fluctuated across the period of the study and that the load was meaningfully associated with changes in the fitness status of players. However, those magnitudes of correlations were small-to-large, suggesting that variations in fitness level cannot be exclusively explained by the accumulated load and loading profile

AVANCES EN EL ESTUDIO DE LA PATOGÉNESIS Y PREVENCIÓN DE LA ENTEROTOXEMIA DE LAS ALPACAS.

The results of our recent research work on enterotoxemia in Peruvian alpacas are presented. Microbiological and molecular analyses found that the majority of the isolates corresponded to Clostridium perfringens and contained the cpa coding gene for α toxin (A genotype) while 0.4% contained both the cpa and cpb genes of the α and β toxins (C genotype). A parallel study revealed that 8.5% of the genotype A isolates also had cpb2, but the cpe (enterotoxin) gene was absent in all cases. These results highly suggest that the exotoxins secreted by C. perfringens are the virulent factors in enterotoxemia, rather than the endogenous enterotoxin. Additionally, an histopathological study of intestinal samples from fatal cases showed that 30.6% had abundant immature structures of Eimeria macusaniensis affecting deep mucosa and cryptic gland epithelia, primarily in the jejune and ileum, suggesting that eimeriosis is likely a triggering or predisposing factor for the development of enterotoxemia. The microbiological studies allowed the design and progressive improvement of an inactivated enterotoxemia vaccine containing primarily the bacterial component plus exotoxins of types A, Aβ2 and C isolated from natural fatal cases of the disease. During six years of field tests in southern Peru, the vaccine has steadily reduced specific neonatal mortality rates due to enterotoxemia from 19.5% (2000, without vaccine) to less than 5% in 2006.Se revisan las investigaciones recientes realizadas por nuestro grupo de investigadores sobre enterotoxemia de las alpacas en el Perú. Estudios microbiológicos y moleculares demostraron que la mayoría de las cepas aisladas fueron de Clostridium perfringens y estas contienen únicamente el gen cpa de la toxina α (C. perfringens genotipo A) y solamente el 0.4% tienen genes cpa y cpb de las toxinas α y β (genotipo C). En análisis paralelo, se encontró que el 8.5% de los genotipos A contenían, adicionalmente, el gen cpb2, pero ninguna cepa tenía el gen cpe de la enterotoxina. Estos resultados evidencian que las exotoxinas secretadas, y no las endotoxinas (cpe), serían los probables factores de virulencia clostridiales en la enterotoxemia de la alpaca. Adicionalmente, en el análisis histopatológico de intestinos infectados, el 30.6% de las muestras presentó abundantes estructuras parasitarias inmaduras correspondientes a Eimeria macusaniensis, afectando la mucosa y epitelio de las glándulas crípticas intestinales, sugiriendo a las infecciones coccidiales como uno de los posibles factores desencadenantes o predisponentes de la enterotoxemia. Estos resultados microbiológicos permitieron diseñar, preparar y mejorar una vacuna convencional inactivada que contiene, mayoritariamente, componentes bacterianos y exotoxinas A, Aβ2 y C aislados de casos fatales de la enfermedad. Desde su introducción en una empresa alpaquera del sur peruano en 2001, la vacuna ha logrado reducir progresivamente los índices de mortalidad por enterotoxemia de 19.5% (2000, sin vacuna) hasta alcanzar tasas menores al 5% en 2006

Concentraciones de inmunoglobulina G sérica en alpacas neonatas muertas por enterotoxemia

The objective of the present study was to determine the levels of serum immunoglobulin G (IgG) in neonatal alpacas (5-23 days of age) killed by enterotoxemia and in animals of similar ages, but clinically healthy. In a first phase, a standard curve of physiological degradation of serum IgG was established from blood sera of six apparently healthy neonates from day 2 to 21 days of age (n=9). In a second phase, serum IgG concentrations were determined in 17 dead neonatal alpacas with lesions compatible with enterotoxemia and in 26 animals of similar ages, apparently healthy. The concentrations of IgG, determined by the Radial Immunodiffusion test, showed that all the animals at 48 hours of birth had adequate concentrations of IgG, while only three of the animals killed by enterotoxemia had IgG levels below the obtained standard curve, although only one of them with levels below 900 mg/dl at 14 days, which could be considered as a partial passive transfer failure. The serum IgG concentrations of the 26 clinically normal animals (2893 mg/dl) and those obtained from animals killed by enterotoxemia (2361 mg/dl) were statistically similar. The risk analysis using the Odds Ratio test (OR: 5.35, CI = 0.50-57.22) indicated that there is no association between adequate levels of IgG and mortality from enterotoxemia in neonatal alpacas.El presente estudio tuvo como objetivo determinar los niveles de Inmunoglobulina G (IgG) sérica en alpacas neonatas (5-23 días de edad) muertas por enterotoxemia y en animales de edades similares, pero clínicamente sanos. En una primera fase se estableció una curva estándar de degradación fisiológica de la IgG sérica a partir de sueros sanguíneos de seis neonatos aparentemente sanos a partir del día 2 y hasta los 21 días de edad (n=9). En una segunda fase se determinaron las concentraciones de IgG sérica en 17 alpacas neonatas muertas con lesiones compatibles con enterotoxemia y en 26 animales, de edades similares, aparentemente sanos. Las concentraciones de IgG, determinadas por la prueba de Inmunodifusión Radial, evidenciaron que todas las crías a las 48 horas del nacimiento presentaron concentraciones adecuadas de IgG, mientras que solo tres de los animales muertos por enterotoxemia tenían niveles de IgG por debajo de la curva estándar de degradación, aunque solo una de ellas con niveles inferiores a 900 mg/dl a los 14 días, lo que podría ser considerado como falla parcial de transferencia pasiva. Las concentraciones de IgG sérica de los 26 animales clínicamente normales (2893 mg/dl) y aquellas obtenidas de animales muertos por enterotoxemia (2361 mg/dl) fueron estadísticamente similares. El análisis de riesgo mediante la prueba de Odds Ratio (OR: 5.35; IC= 0.50-57.22) indicó que no existe asociación entre niveles adecuados de IgG y la mortalidad por enterotoxemia en alpacas neonatas

Comparison of two modes of vitamin B12 supplementation on neuroconduction and cognitive function among older people living in Santiago, Chile: a cluster randomized controlled trial. a study protocol [ISRCTN 02694183]

BACKGROUND: Older people have a high risk of vitamin B12 deficiency; this can lead to varying degrees of cognitive and neurological impairment. CBL deficiency may present as macrocytic anemia, subacute combined degeneration of the spinal cord, or as neuropathy, but is often asymptomatic in older people. Less is known about subclinical vitamin B12 deficiency and concurrent neuroconduction and cognitive impairment. A Programme of Complementary Feeding for the Older Population (PACAM) in Chile delivers 2 complementary fortified foods that provide approximately 1.4 μg/day of vitamin B12 (2.4 μg/day elderly RDA). The aim of the present study is to assess whether supplementation with vitamin B12 will improve neuroconduction and cognitive function in older people who have biochemical evidence of vitamin B12 insufficiency in the absence of clinical deficiency. METHODS: We designed a cluster double-blind placebo-controlled trial involving community dwelling people aged 70-79 living in Santiago, Chile. We randomized 15 clusters (health centers) involving 300 people (20 per cluster). Each cluster will be randomly assigned to one of three arms: a) a 1 mg vitamin B12 pill taken daily and a routine PACAM food; b) a placebo pill and the milk-PACAM food fortified to provide 1 mg of vitamin B12; c) the routine PACAM food and a placebo pill.The study has been designed as an 18 month follow up period. The primary outcomes assessed at baseline, 4, 9 and 18 months will be: serum levels of vitamin B12, neuroconduction and cognitive function. CONCLUSIONS: In view of the high prevalence of vitamin B12 deficiency in later life, the present study has potential public health interest because since it will measure the impact of the existing program of complementary feeding as compared to two options that provide higher vitamin B12 intakes that might potentially may contribute in preserving neurophysiologic and cognitive function and thus improve quality of life for older people in Chile. TRIAL REGISTRATION: ISRCTN: ISRCTN02694183

Towards precision medicine: defining and characterizing adipose tissue dysfunction to identify early immunometabolic risk in symptom-free adults from the GEMM family study

Interactions between macrophages and adipocytes are early molecular factors influencing adipose tissue (AT) dysfunction, resulting in high leptin, low adiponectin circulating levels and low-grade metaflammation, leading to insulin resistance (IR) with increased cardiovascular risk. We report the characterization of AT dysfunction through measurements of the adiponectin/leptin ratio (ALR), the adipo-insulin resistance index (Adipo-IRi), fasting/postprandial (F/P) immunometabolic phenotyping and direct F/P differential gene expression in AT biopsies obtained from symptom-free adults from the GEMM family study. AT dysfunction was evaluated through associations of the ALR with F/P insulin-glucose axis, lipid-lipoprotein metabolism, and inflammatory markers. A relevant pattern of negative associations between decreased ALR and markers of systemic low-grade metaflammation, HOMA, and postprandial cardiovascular risk hyperinsulinemic, triglyceride and GLP-1 curves was found. We also analysed their plasma non-coding microRNAs and shotgun lipidomics profiles finding trends that may reflect a pattern of adipose tissue dysfunction in the fed and fasted state. Direct gene differential expression data showed initial patterns of AT molecular signatures of key immunometabolic genes involved in AT expansion, angiogenic remodelling and immune cell migration. These data reinforce the central, early role of AT dysfunction at the molecular and systemic level in the pathogenesis of IR and immunometabolic disorders