Location also matters: The oxidative response of the intertidal purple mussel Perumytilus purpuratus during tidal cycle.

For sessile intertidal organisms, periods of low tide impose both cellular and physiological challenges that can determine bathymetric distribution. To understand how intertidal location influences the cellular response of the bivalve Perumytilus purpuratus during the tidal cycle (immersion-emersion-immersion), specimens from the upper intertidal (UI) and lower intertidal (LI) of bathymetric distribution were sampled every 2 h over a 10-h period during a summer tidal cycle. Parallelly, organisms from the UI and LI were reciprocally transplanted and sampled throughout the same tidal cycle. Levels of oxidative damage (lipid peroxidation and protein carbonyls) as well as total antioxidant capacity and total carotenoids were evaluated as cellular responses to variations in environmental conditions throughout the tidal cycle. The results indicate that both the location in the intertidal zone (UI/LI), the level of aerial exposure, and the interaction of both factors are determinants of oxidative levels and total antioxidant capacity of P. purpuratus. Although oxidative damage levels are triggered during the low tide period (aerial exposure), it is the UI specimens that induce higher levels of lipid peroxidation compared to those from the LI, which is consistent with the elevated levels of total antioxidant capacity. On the other hand, organisms from the LI transplanted to the UI increase the levels of lipid peroxidation but not the levels of protein carbonyls, a situation that is also reflected in higher levels of antioxidant response and total carotenoids than those from the UI transplanted to the LI. The bathymetric distribution of P. purpuratus in the intertidal zone implies differentiated responses between organisms of the lower and upper limits, influenced by their life history. A high phenotypic plasticity allows this mussel to adjust its metabolism to respond to abrupt changes in the surrounding environmental conditions.

Upwelling as a stressor event during embryonic development: Consequences for encapsulated and early juvenile stages of the marine gastropod Acanthina monodon.

Upwelling phenomena alter the physical and chemical parameters of the sea's subsurface waters, producing low levels of temperature, pH and dissolved oxygen, which can seriously impact the early developmental stages of marine organisms. To understand how upwelling can affect the encapsulated development of the gastropod Acanthina monodon, capsules containing embryos at different stages of development (initial, intermediate and advanced) were exposed to upwelling conditions (pH = 7.6; O2 = 3 mg L-1; T° = 9 °C) for a period of 7 days. Effects of treatment were determined by estimating parameters such as time to hatching, number of hatchlings per capsule, percentage of individuals with incomplete development, and shell parameters such as shell shape and size, shell strength, and the percentage of the organic/inorganic content. We found no significant impacts on hatching time, number of hatchlings per capsule, or percentage of incomplete development in either the presence or absence of upwelling, regardless of developmental stage. On the other hand, latent effects on encapsulated stages of A. monodon were detected in embryos that had been exposed to upwelling stress in the initial embryonic stage. The juveniles from this treatment hatched at smaller sizes and with higher organic content in their shells, resulting in a higher resistance to cracking 30 days after hatching, due to greater elasticity. Geometric morphometric analysis showed that exposure to upwelling condition induced a change in the morphology of shell growth in all post-hatching juveniles (0-30 days), regardless of embryonic developmental stage at the time of exposure. Thus, more elongated shells (siphonal canal and posterior region) and more globular shells were observed in newly hatched juveniles that had been exposed to the upwelling condition. The neutral or even positive upwelling exposure results suggests that exposure to upwelling events during the encapsulated embryonic phase of A. monodon development might not have major impacts on the future juvenile stages. However, this should be taken with caution in consideration of the increased frequency and intensity of upwelling events predicted for the coming decades.

Embryonic encapsulated development of the gastropod Acanthina monodon is impacted by future environmental changes of temperature and pCO2.

Egg capsules of the gastropod Acanthina monodon were maintained during the entire period of encapsulated development at three temperatures (10, 15, 20 °C) and two pCO2 levels (400, 1200 µatm). Embryos per capsule, size at hatching, time to hatching, embryonic metabolic rates, and the resistance of juveniles to shell breakage were quantified. No embryos maintained at 20 °C developed to hatching. The combination of temperature and pCO2 levels had synergistic effects on hatching time and developmental success, antagonistic effects on number of hatchlings per capsule, resistance to juvenile shell cracking and metabolism, and additive effect on hatching size. Juveniles hatched significantly sooner at 15 °C, independent of the pCO2 level that they had been exposed to, while individuals hatched at significantly smaller sizes if they had been held under 15 °C/1200 µatm rather than at 10 °C/low pCO2. Embryos held at the higher pCO2 had a significantly greater percentage of abnormalities. For capsules maintained at low pCO2 and 15 °C, emerging juveniles had less resistance to shell breakage. Embryonic metabolism was significantly higher at 15 °C than at 10 °C, independent of pCO2 level. The lower metabolism occurred in embryos maintained at the higher pCO2 level. Thus, in this study, temperature was the factor that had the greatest effect on the encapsulated development of A. monodon, increasing the metabolism of the embryos and consequently accelerating development, which was expressed in a shorter intracapsular development time, but with smaller individuals at hatching and a lower resistance of their shells to breakage. On the other hand, the high pCO2 level suppressed metabolism, prolonged intracapsular development, and promoted more incomplete development of the embryos. However, the combination of the two factors can mitigate--to some extent--the adverse effects of both incomplete development and lower resistance to shell breakage.

UV-R mitigation strategies in encapsulated embryos of the intertidal gastropod Acanthina monodon: A way to compensate for lack of parental care.

Intracapsular embryonic development in the intertidal zone exposes embryos to various stress sources characteristic of this environment, including UV-R. They require defensive mechanisms to mitigate its adverse effects. The presence of total carotenoids (TC), and mycosporine-like amino acids (MAAs) was studied in adults, in encapsulated embryos, and in the egg capsule walls of the intertidal gastropod Acanthina monodon. Oxygen consumption rates (OCR) were determined in encapsulated and excapsulated embryos exposed to photosynthetically active radiation (PAR) and PAR + UV-A + UV-B to understand if the capsule wall is a protective structure for encapsulated embryos. The results showed the presence of TC in adult pedal and gonad tissues, and in all encapsulated stages. MAAs were not detected. The physical structure of the capsule wall retained most wavelengths, being particularly efficient in the UV-B range. Excapsulated embryos exposed to PAR + UV-A + UV-B radiation increased its OCR compared to encapsulated embryos, indicating the protective character of the capsule wall.

Relationship between over-crowding within egg capsules of the marine gastropod Acanthina monodon and prospects for juvenile success.

Encapsulated development with extraembryonic yolk may lead to competition for nutrients within egg capsules. In this research, different degrees of competition among embryos in subtidal egg capsules of Acanthina monodon resulted in considerable differences in hatching size. For newly hatched juveniles, individuals hatching from less crowded egg capsules showed better survival, larger SL, higher rates of oxygen consumption, and higher rates of food consumption. However, by 28 days after hatching, the largest surviving juveniles were the best-performing individuals, regardless of the initial embryo density within the capsules. In summary, more crowded egg capsules resulted in poorer survival. These findings may help to explain the variability seen in juvenile success in some field populations; much of that variation may reflect stressful experiences that the new recruits have had during the early stages of their encapsulated development.

Respiratory and desiccation constraints during encapsulated intertidal development of the marine gastropod Acanthina monodon.

Acanthina monodon commonly deposits its egg capsules in the intertidal zone. Capsule aerial exposure during low-tide can impact oxygen consumption rates (OCR) of embryos and intracapsular oxygen availability, and expose embryos to desiccation. OCR increased as embryonic development progressed, and was greater when capsules were submerged in seawater than when exposed to air. Oxygen available within the capsule was always less than that available in the immediate external environment, whether capsules were immersed or exposed. The highest internal oxygen concentrations were recorded during periods of air exposure for embryos in more advanced development stages. When exposed to air, capsules lost water the fastest when they contained early embryos, and suffered the highest mortalities following exposure. Collectively, these data suggest that, although encapsulation helps the embryos to develop across wildly fluctuating environmental conditions, the amount of stress the embryos experience will vary depending on their exact positioning within the intertidal zone.

Are embryonic developing modes determinant in the acquisition and levels of photoprotective compounds in slipper limpets of the Crepipatella genus?

The type of embryonic development (mixed and direct) and its influence on the accumulation and translocation of photoprotective compounds from the mother to the encapsulated embryo was studied in the intertidal gastropods Crepipatella peruviana and Crepipatella dilatata during their reproductive peak. HPLC/MS was used to determine type and levels of sunscreen compounds (total carotenoids; TC/and mycosporine-like amino acid; MAA) in brooding females, capsule walls and early and pre-hatching embryos of both species. Photoprotective compounds were only quantified in nurse eggs of C. dilatata. Our results indicate that females of both species can accumulate TC and MAA at different levels, and they are able to transfer them selectively to capsule walls, embryos and nurse eggs. Palythine-serine (MW=244Da; λmax=320nm) and MAA-330 (MW=234Da; λmax=330nm) constitute total MAA pool in brooding females, whereas brooded embryos incorporate palythine (MW=244Da; λmax=320nm) to the MAA pool. Although TC was transferred from the mother to the embryo through the yolk in both species, MAA trespass showed differences. Females of C. peruviana transfer MAA to their embryos through the embryonic yolk; C. dilatata can transfer MAA only through their nurse eggs, which are consumed by embryos during the terminal stages of intracapsular development. Differences between mixed and direct embryonic development, as well as environmental UV-R levels, which the recently hatched larvae and juveniles of C. peruviana and C. dilatata are exposed to, would determine levels of sunscreen compounds in each species. Higher TC and MAA levels in pre-hatching larvae of C. peruviana compared to C. dilatata, indicate a necessity of C. peruviana for protection against UV-R radiation during approximately 15days when their veliger larvae remain in the water column before metamorphosis is complete. Conversely, low photoprotective levels in pre-hatching juveniles of C. dilatata could be related to low UV-R exposure levels due to the direct incorporation to the benthos and the presence of a protective shell.

Variabilidad en la asistencia hospitalaria. El caso de la rehabilitación respiratoria en Colombia / Variability in hospital care. Pulmonary rehabilitation in Colombia

Objetivo: Describir las variaciones en la asistencia hospitalaria de los programas de rehabilitación respiratoria (RR) de pacientes con enfermedad pulmonar obstructiva crónica (EPOC) en un colectivo de fisioterapeutas colombianos. Materiales y métodos: Estudio observacional con diseño transversal en 250 profesionales de fisioterapia de Colombia. Se aplicó una encuesta para identificar las variaciones en los procedimientos, los componentes, la estructura y las características de los programas de RR, según las «directrices del manejo fisioterapéutico en paciente con enfermedad pulmonar con respiración espontánea» publicadas por la British Thoracic Society (BTS). Adicionalmente, se reunieron datos de información sociodemográfica. Resultados: La tasa de respuesta fue del 89,6% (n = 224). En el momento de la encuesta, el (92%) trabajaba en su mayor tiempo en programas de RR. Respecto a las variaciones en la práctica clínica, un elevado porcentaje de profesionales respondieron «realizar siempre» actividades con bajo nivel de evidencia (grados C y D), como técnicas de respiración diafragmática (48,7%), respiración con labios fruncidos durante el esfuerzo (62,9%) y técnicas de control de la respiración (37,5%). También se observó la inclusión de intervenciones con mayor grado de recomendación (grado A), como entrenamiento muscular periférico (42,9%), entrenamiento de músculos respiratorios (52,2%), educación y/o asesoramiento de la enfermedad (56,7%), uso de ventilación con presión positiva intermitente (31,7%), y aplicación de terapia de oxígeno, tanto en la fase aguda como domiciliaria (40,6%). Conclusiones: El estudio confirma importantes variaciones en los componentes terapéuticos asistenciales de la RR planteadas por la BTS para pacientes con EPOC

Objective: To describe variations in the management of pulmonary rehabilitation (PR) programs in patients with Chronic Obstructive Pulmonary Disease (COPD) by Colombian physiotherapists. Materials and method: We conducted a cross-sectional, observational study with 250 Colombian physiotherapists who routinely work with COPD patients from Colombia. A survey was made to identify variations in procedures, components, structure and characteristics of PR programs according to the «Guidelines for the physiotherapy management in patients with pulmonary disease with spontaneously breathing» published by British Thoracic Society Physiotherapy (BTS). Additionally, socio-demographic data were collected. Results: The response rate was 89.6% (n = 224). At the time of the survey, 92% of those surveied spent most of their time working in rehabilitation programs. Regarding variations in the clinical practice, the physiotherapists mainly answered «always perform» activities with low level of evidence (Grade C and D) such as diaphragmatic breathing techniques (48.7%), pursed-lip breathing during exercise (62.9%) and techniques of breath control (37.5%). A high percentage of professionals also included interventions with a higher degree of recommendation (Grade A) such as peripheral muscle training (42.9%), respiratory muscle training (52.2%), education or counseling disease (56.7%), use of intermittent positive pressure ventilation (31.7%), and application of oxygen (40.6%). Conclusions: The study confirms that there are important variations in therapeutic care components of the PR proposed by the BTS for patients with Chronic Obstructive Pulmonary Disease

Effects of ultraviolet radiation on an intertidal trematode parasite: an assessment of damage and protection.

Trematode parasites are integral components of intertidal ecosystems which experience high levels of ultraviolet radiation. Although these parasites mostly live within hosts, their life cycle involves free-living larval transmission stages such as cercariae which are directly exposed to ambient conditions. UV has previously been shown to considerably reduce the survival of cercariae. Here, we investigated potential mechanisms of protection and damage related to UV in the intertidal trematode Maritrema novaezealandensis. Firstly, the presence of sunscreen compounds (i.e. mycosporine-like amino acids) was quantified in the parasite tissue producing cercariae within a snail host, as well as in the free-swimming cercariae themselves. Secondly, levels of oxidative stress in cercariae after exposure to UV were investigated (i.e.protein carbonyls, catalase and superoxide dismutase). Thirdly, the DNA damage (i.e. cyclobutane­pyrimidine dimers) was compared between cercariae exposed and not exposed to UV. Lastly, functional aspects(survival and infectivity) of cercariae were assessed, comparing cercariae under light conditions versus dark after exposure to UV. We confirmed the presence of my cosporine-like amino acids in cercariae-producing tissue from within snail hosts, but were unable to do so in cercariae directly. Results further suggested that exposure to UV induced high levels of oxidative stress in cercariae which was accompanied by a reduction in the levels of protective antioxidant enzymes present. We also identified higher levels of DNA damage in cercariae exposed to UV, compared with those not exposed. Moreover, no clear effect of light condition was found on survival and infectivity of cercariae after exposure to UV. We concluded that cercariae are highly susceptible to UV damage and that they have very little scope for protection against or repair of UV-induced damage.

Characterisation of strains of infectious bronchitis virus isolated in Chile.

Nine isolates of infectious bronchitis (IB)-like viruses were made from 23 flocks (broilers or layers) in Chile experiencing the types of disease problems commonly associated with IBV. Their identity as IB viruses was confirmed. The histological changes they caused in tracheal organ cultures (OC) are described. Serum neutralisation tests performed in embryonated eggs (alpha-method) suggested that four of the isolates were serologically related to the Massachusetts (Mass) serotype of IBV and one to Connecticut. The four other strains were examined further by a serum neutralisation test in OC (ss-method). One was found to be of the Mass serotype but the others were found to be unrelated antigenically to a wide range of IBV serotypes isolated in many countries over a number of years. One of these three strains and the Mass strain, when given intranasally to 8-day-old specified pathogen free chickens together with pathogenic serotypes of E.coli, caused some mortality and considerable morbidity. The H120 vaccine strain was found not to protect completely against challenge with these four strains 21 days later.

[Experimental vaccines against avian inclusion body hepatitis. I. Serological evaluation]. / Vacunas experimentales contra hepatitis A cuerpo de inclusión aviar. I. Valoración serológica.

Two Inclusion Body Hepatitis (IBH) vaccines (Vac-19 and Vac-28) were elaborated from serotypes 4, 10 and 11, isolated from country field outbreaks. The strains were attenuated by passages in embryonating SPF chicken eggs (16 and 22 passages for both vaccines) and in kidney cell culture (3 and 6 passages respectively). Four experimental groups (G) were used: G I and G II of 8 chickens each, vaccinated orally (4,4 TCID50/ml) with Vac-19 and Vac-28 respectively, at the third week and re-vaccinated 35 days later with the same dose and method. Groups G III and G IV with 5 chickens each were the positive and negative controls, both group were not vaccinated, the first only challenged wit a pool of the 4, 10 and 11 serotypes on 45 days old and the last was not challenged. Prior and afterward, vaccination and challenge, clinical and serological exams were performed. The immune response was determined trough Agar Gel Precipitin (AGP) and serum neutralization (SN) test in chicken kidney cells (CKC) using the beta method (200 TCID50/50 microliters). Positive response (100%) was detected from day 43 onward in the vaccinated group, to AGP test. This situation occurred on day 55 for the challenged group (G III). Geometric mean titres (GMT) were, prior to challenge, 1.194 for G I and 23.525 for G II. After challenge, GMT were 7.760 and > or = 305.736 respectively. The GMT for G III was 640. No clinical signs or mortality were observed during the experiment. The conclusion is that only Vac-28 provided a partial protection to the fowl, measured through the lesions observed.

[Parasitism in fishes and human riverside communities of the Huillinco and Natri lakes (Great Island of Chiloé), Chile]. / Parasitismo en peces y comunidades humanas ribereñas de los lagos Huillinco y Natri (Isla Grande de Chiloé), Chile.

In April and December 1989, 35 fish from Lake Huillinco (42 degrees 48'S, 74 degrees 02'W) and 36 fish from Lake Natri (42 degrees 48'S, 73 degrees 50'W), in the Great Island of Chiloé (Chile) were examined. Coprological samples from 159 persons, 17 dogs, 19 pigs and 4 cats from around both lakes were examined for Diphyllobothrium spp. infection. In the Lake Huillinco the following helminths of fishes were determined: Contracaecum sp. and Hysterothylacium sp. in Salmo trutta, Cauque mauleanum and Eleginops maclovinus; Dichelyne (Cucullanellus) dichelyneformis in S. trutta and E. maclovinus and Scolex pleuronectis in S. trutta. One specimen of Mugil cephalus did not show helminth parasites. Prevalence of infection were greater for Contracaecum sp. in S. trutta (75.0%) and C. mauleanum (76.0%); and Hysterothylacium sp. in E. maclovinus (75.0%). Mean intensity was higher for D. (C.) dichelyneformis in E. maclovinus. Contracaecum sp. in S. trutta, Oncorhynchus mykiss, Oncorhynchus kisutch and Galaxias maculatus; Acanthocephalus sp. in S. trutta and G. maculatus, S. pleuronectis in O. mykiss and Cystidicoloides sp. in G. maculatus were determined at Lake Natri. Prevalence and intensity of infection were higher for Contracaecum sp. in S. trutta and O. kisutch. Infection by Diphyllobothrium sp. was determined in one domestic cat. Prevalence of infection by intestinal protozoan and helminths in human population only showed significative differences for Ascaris lumbricoides and Trichuris trichiura that were higher in the Lake Huillinco. Importance of natural infection by helminth parasites for fish in cultured condition and possible mechanisms of infections in relation to the diet of fishes are discussed.

Serum changes and histologic liver lesions due to experimental ingestion of ragwort (Senecio erraticus) in sheep.

Eight 1-year-old Romney-Marsh female sheep were used to test the effect of Senecio erraticus (ragwort). Animals were fed for 60 days on a standard diet incorporating 20% dried ground ragwort. Morphology of the livers was studied by serial biopsy and function by changes in plasma bilirubin concentration, the half-life of bromosulphthalein (BSP), and by the activity of glutamate dehydrogenasa (GD) in plasma during and after Senecio ingestion. All sheep had an increase in bilirubin concentration and GD activity in their plasma and in BSP half-life. These values returned to normal 30 days after ragwort ingestion ceased. Histological studies showed Kupffer cell hypertrophy, fatty degeneration and megalocytosis. These lesions almost disappeared 60 days after stopping Senecio ingestion.