Effect of ß-glucans on rainbow trout (Oncorhynchus mykiss) IgM+ B cells.

ß-glucans are carbohydrates present in the cell wall of many fungi, which are often used as immunostimulants in feeds for farmed species. Their capacity to activate innate immune responses directly acting on innate cell populations has been widely documented in fish. However, whether they can affect the functionality of adaptive immune cells has been scarcely explored. In this context, in the current work, we have determined the effects of ß-glucans on rainbow trout blood IgM+ B cells in the presence or absence of 2,4,6-trinitrophenyl hapten conjugated to lipopolysaccharide (TNP-LPS), a model antigen. For this, rainbow trout peripheral blood leukocytes were incubated with different doses of ß-glucans or media alone in the presence or absence of TNP-LPS for 48 h. The size, levels of expression of surface MHC II, antigen processing and phagocytic capacities and proliferation of IgM+ B cells were then studied by flow cytometry. The number of IgM-secreting cells in the cultures was also estimated by ELISpot. ß-glucans significantly decreased the levels of surface MHC II expression and the antigen processing capacities of these cells, especially in the presence of TNP-LPS, while they increased their phagocytic activity. On their own, ß-glucans slightly activated the proliferation of IgM+ B cells but reduced that induced by TNP-LPS. In contrast, ß-glucans significantly increased the number of cells secreting IgM in the cultures. This effect of ß-glucans on the IgM-secreting capacity of B cells was also confirmed through a feeding experiment, in which the IgM-secreting capacity of blood leukocytes obtained from fish fed a ß-glucan-supplemented diet for one month was compared to that of leukocytes obtained from fish fed a control diet. Altogether, these findings contribute to increase our knowledge regarding the effects of ß-glucans on fish adaptive responses.

Olfactory bulb plasticity ensures proper olfaction after severe impairment in postnatal neurogenesis.

The olfactory bulb (OB) neurons establish a complex network that ensures the correct processing of the olfactory inputs. Moreover, the OB presents a lifelong addition of new neurons into its existing circuitry. This neurogenesis is considered essential for the OB function. However, its functional impact on physiology and behavior is still unclear. Here, we investigate the mechanisms of OB plasticity that underlie bulbar physiology in relation to severe damage of neurogenesis. The neurogenesis of young mice was altered by ionizing radiation. Afterwards, both multi-channel olfactometry and electrophysiological studies were performed. Furthermore, neurogenesis and differentiation of the newly formed cells were assessed using bromodeoxyuridine labeling combined with a wide battery of neuronal markers. Our results demonstrate a reduction in both neurogenesis and volume of the OB in irradiated animals. The number of neuroblasts reaching the OB was reduced and their differentiation rate into interneurons selectively changed; some populations were noticeably affected whereas others remained preserved. Surprisingly, both olfactory detection and discrimination as well as electrophysiology presented almost no alterations in irradiated mice. Our findings suggest that after damaging postnatal neurogenesis, the neurochemical fate of some interneurons changes within a new biological scenario, while maintaining homeostasis and olfaction.

Real-time RT-PCR detection of betanodavirus in naturally and experimentally infected fish from Spain.

Infections with betanodavirus affect a wide range of wild and farmed fish species throughout the world, mostly from the marine environment. The aim of this work was to develop and validate real-time RT-PCR assays for sensitive and specific detection of nodavirus in diseased or carrier fish. The new detection assay was used to study the transmission and development of nodavirus infection in juvenile sea bass, Dicentrarchus labrax (L.), challenged by different routes, and also to screen for nodavirus in various farmed fish species. On average, the sensitivity was 10-100 times higher than a standard RT-PCR, and the assay was able to detect asymptomatic carrier fish that otherwise could have been classified as free of infection. Clinical signs of nodavirus infection were reproduced in fish infected following bath exposure or intramuscular injection, demonstrating horizontal transmission of the disease. Nodavirus was always detected in the brain of diseased fish but also in many recovered fish. The new assay enables us to confirm the presence of the virus at an early phase in the production cycle and may represent a useful tool to prevent or slow down the spread of nodavirus to new locations.

Cardicola aurata sp. n. (Digenea: Sanguinicolidae) from Mediterranean Sparus aurata L. (Teleostei: Sparidae) and its unexpected phylogenetic relationship with Paradeontacylix McIntosh, 1934.

A new sanguinicolid trematode, Cardicola aurata sp. n., is described from gilthead seabream Sparus aurata L., from off the Spanish Mediterranean coast. The morphology of C. aurata sp. n. generally agrees with the diagnosis of the genus, however, in contrast to all other reported Cardicola spp. the male pore is located sub-medially at the posterior end of the body instead of sinistrally before the posterior end of the body. Based on a comparison of the morphology as well as partial 28S and ITS2 rDNA sequence data from the present species with that from closely related species, it was decided to emend the diagnosis of Cardicola rather than create a new genus, as the aberrant position of the male pore is likely to be an autapomorphy. The phylogenetic analyses revealed a close relationship between Cardicola and Paradeontacylix, two genera with considerable morphological differences; C. aurata sp. n. occupies a position intermediate to these genera. Thus, a morphological comparison of Cardicola, Paradeontacylix and Braya, a genus which is morphologically similar to Cardicola but clusters basal to the Cardicola/Paradeontacylix clade, was conducted. The results of this comparison showed that despite large differences with regard to body shape, the organisation of the internal organs is very similar in species of Cardicola and Paradeontacylix. The synopsis of morphological data and molecular phylogeny allows for interpretations regarding the importance of different morphological features for the phylogenetic inference of the Sanguinicolidae.

Controllable versus uncontrollable stressors bi-directionally modulate conditioned but not innate fear.

Fear conditioning and fear extinction play key roles in the development and treatment of anxiety-related disorders, yet there is little information concerning experiential variables that modulate these processes. Here we examined the impact of exposure to a stressor in a different environment on subsequent fear conditioning and extinction, and whether the degree of behavioral control that the subject has over the stressor is of importance. Rats received a session of either escapable (controllable) tail shock (ES), yoked inescapable (uncontrollable) tail shock (IS), or control treatment (home cage, HC) 7 days before fear conditioning in which a tone and foot shock were paired. Conditioning was measured 24 h later. In a second experiment rats received ES, IS or HC 24 h after contextual fear conditioning. Extinction then occurred every day beginning 7 days later until a criterion was reached. Spontaneous recovery of fear was assessed 14 days after extinction. IS potentiated fear conditioning when given before fear conditioning, and potentiated fear responding during extinction when given after conditioning. Importantly, ES potently interfered with later fear conditioning, decreased fear responding during fear extinction, and prevented spontaneous recovery of fear. Additionally, we examined if the activation of the ventral medial prefrontal cortex (mPFCv) by ES is critical for the protective effects of ES on later fear conditioning. Inactivation of the mPFCv with muscimol at the time of the initial experience with control prevented ES-induced reductions in later contextual and auditory fear conditioning. Finally, we explored if the protective effects of ES extended to an unconditioned fear stimulus, ferret odor. Unlike conditioned fear, prior ES increased the fear response to ferret odor to the same degree as did IS.

First description of non-motile Yersinia ruckeri serovar I strains causing disease in rainbow trout, Oncorhynchus mykiss (Walbaum), cultured in Spain.

Yersinia ruckeri, the causal agent of enteric redmouth (ERM) disease, was isolated from epizootics that occurred in different Spanish rainbow trout, Oncorhynchus mykiss (Walbaum), farms in which vaccination against ERM had been performed. In all episodes, the most pronounced clinical signs exhibited by affected fish were severe haemorrhages in the mouth, eyes and around the vent. The isolates were identified as Y. ruckeri serovar I by 16S rRNA sequencing together with serological tests. They lacked motility and lipase activity and thus belonged to biotype 2, and were highly virulent for juvenile rainbow trout, both by intraperitoneal injection (from 3.1 x 10(2) to 6.3 x 10(3) cfu per fish) and bath challenge (5.1-7.3 x 10(6) cfu mL(-1)). This is the first description of Y. ruckeri serovar I biotype 2 causing disease in cultured trout in Spain vaccinated with commercial ERM vaccines. The occurrence of this emergent pathogen in Spanish continental aquaculture from its first isolation in 2001 to date is also documented.

Pathology of Edwardsiella tarda infection in turbot, Scophthalmus maximus (L.).

Macroscopic and histopathological changes in cultured turbot, Scophthalmus maximus (L.), in Spain caused by infection with Edwardsiella tarda are described. Eye tumefaction, inflammation, haemorrhages, ascites and the presence of a purulent fluid were the main macroscopic lesions observed. Histopathological lesions were found in the kidney, spleen and liver. In the kidney and spleen these were characterized by a severe apostematous inflammatory reaction, with a large number of abscesses. The liver was affected to a lesser degree and only some phagocytes loaded with bacteria were observed. Ultrastructural observations indicated that macrophages were the main cell type implicated in the inflammatory response. Most of the bacteria observed within the phagocyte cytoplasm showed no degenerative changes and some were dividing. Degenerative changes observed in macrophages indicate their failure in preventing the infection.

Evaluation of a CD61 MoAb method for enumeration of platelets in thrombocytopenic patients and its impact on the transfusion decision-making process.

BACKGROUND: Almost all automated hematology cell analyzers use methods based on either the impedance (PLTi) or the optical (PLTo) properties of the cells for performing platelet counts. To improve the accuracy of platelet counts in peripheral blood (PB), the use of CD61 (GPIIIa) MoAbs (ImmunoPLT method) has recently been introduced in an automated hematology blood-analyzer system (Cell-Dyn 4000, Abbott Diagnostics). STUDY DESIGN AND METHODS: A comparative evaluation was made of the accuracy and precision of the three methods currently available in the Cell-Dyn 4000 automated hematology cell analyzer for counting the number of platelets per microliter of PB in a total of 47 patients with chemotherapy-induced thrombocytopenia. A flow cytometric PB platelet count was also performed in parallel and used as an external reference. RESULTS: PB platelet counts showed a good correlation among the PLTo, CD61-ImmunoPLT, and flow cytometric methods. In contrast, the PLTi procedure usually provided an overestimation of the number of platelets per microliter. Although a good correlation was observed between the flow cytometric reference method and both the ImmunoPLT and PLTo methods, the highest degree of agreement was found for the ImmunoPLT techniques (94% vs. 67%). A comparative analysis of the PLTo and CD61-ImmunoPLT methods with regard to their value for predicting platelet transfusion needs on the basis of specific flow cytometric platelet count thresholds showed a good correlation when the cutoff level of 10,000 platelets per microL was used. In contrast, at the threshold of 20,000 platelets per microL, slight differences were observed between the PLTo and CD61-ImmunoPLT procedures for predicting transfusion needs. CONCLUSION: Such results indicate that, if the CD61-ImmunoPLT method is used in the platelet transfusion decision-making process, unnecessary platelet transfusions could be avoided in up to 17.5 percent of persons with a PLTo count of <20,000 platelets per microL.

Histopathology of cultured sea bream Sparus aurata infected with sanguinicolid trematodes.

The present study is the first report of a sanguinicolid infection affecting sea bream Sparus aurata cultured in net cages in the NE of Spain. The disease was associated with trickling mortalities during the cold season (1999 and 2000). Examination of gill wet mounts of the affected population revealed that sanguinicolid infection was present in 82.6 and 100% of the fish sampled in 1999 and 2000, respectively. Adult flukes, which were located in the kidney, were tentatively identified as members of the family Sanguinicolidae, subfamily Cardicolinae. Eggs and miracidia were found in the gill vascular structures. The inflammatory response triggered by the parasites was moderate and the lesions caused by either eggs and miracidia in the gills or adult flukes in the kidney were not extremely severe, possibly because of the moderate intensity of the parasitosis. Histological observations of sanguinicolid infected sea bream presented here are compared with those reported in other fish species. The role played on sea bream morbility and mortality by other factors (occurrence of a simultaneous moderate monogenean infection, immunological impairement related to low water temperatures) is discussed.

Myxidium leei (Myxozoa) infections in aquarium-reared Mediterranean fish species.

An episode of parasitic enteritis causing trickling mortalities at an exhibition aquarium reproducing Mediterranean ecosystems was found to be caused by the myxozoan parasite Myxidium leei Diamant, Lom & Dykova 1994. The myxozoan was recorded in 25 different fish species belonging to 16 Genera, 10 Families and 4 Orders. It was mainly detected in the intestine of affected fish, and was responsible for severe chronic enteritis. The parasite was probably introduced into the facilities with infected wild fish, and transmitted directly from fish to fish by cohabitation, transfer of infected material and necrophagia. Fish belonging to the Families Labridae and Blenniidae appeared as most susceptible, and the incidence of infections in members of the Sparidae was low. This study significantly widens the host spectrum for this virulent parasite and now includes many ubiquitous coastal Mediterranean species. Wild fish may have a significant role in the transmission of myxidiosis of cultured sparid fish.

[Obesity, arterial hypertension, metabolic disorders, and polycystic ovary syndrome]. / Obesidad, hipertensión arterial, alteraciones metabólicas y el síndrome de ovarios poliquísticos (SOP).

OBJECTIVE: To determine the prevalence of central and general obesity, arterial hypertension, and alterations in cholesterol, triglycerids, glycaemia and uric acid levels in patients with PCOS and to compare them with women who do not present this syndrome. To establish the possible association of this with the PCOS. MATERIALS AND METHODS: This is a transversal analytic study. It includes fifty-five cases of PCOS and fifty-five healthy controls from gynecologic and familiar medicine consultation during the period from May 2nd to November 30th, 1999. It excludes patients with the Cushing disease, congenital suprarenal hyperplasia, pregnant women or women under hormonal treatments until three months prior the beginning of the study. The diagnostic of PCOS was established when the patient presents three of the following characteristics: menstrual disturbances, hirsutism, a LH/FSH relation > or = than 1.5 and ultrasound with polycystic ovaries. Their weight and height was taken in order to calculate the body mass index (BMI) and the waist/hip index (WHI) Blood samples in fasting where obtained to determine the FSH, LH, cholesterol, triglycerids, glycaemia and uric acid levels. Descriptive statistics where applied as well as the SPSS software along with the Saphiro-Francia test, the Mann Whitney test and logistic regression. RESULTS: The age of the patients with PCOS varied between 18 and 39 years old, average 26.6 years old and that of the controls from 19 to 39 years old, average 28.2 years old, (p = 0.165). The prevalence of general obesity (BMI > or = 30) was of 50.9% in the group of PCOS and of 18.2% in the group of controls. The systolic hypertension (BP > or = 140 mm Hg) was of 9% against 7.3%. A diastolic hypertension (BP > or = 90 mm Hg) of 25.5% against 7.3%. A hypercholesterolemia (> or = 220 mg/dL) of 30.9% against 10.9%. A hypertriglyceridemia (> or = 160 mg/dL) of 30.9% against 16.4%. A hyperglycaemia(> or = 115 mg/dL) of 5.5% against 3.6% and a hyperuricemia(> or = 6.5 mg/L) of 23.6% against 3.6%. The OR calculated through cross products showed that patients with PCOS have the possibility of presenting obesity, hypercholesterolemia, hypertriglyceridemia, hyperuricemia and diastolic hypertension, 4.6, 3.7, 2.3, 8.2 and 4.4 times more than the controls. The comparison of the average of the variables of both groups showed significant differences (p < 0.5) for the BMI, WHI, systolic BP, cholesterol and uric acid. The multiple logistic regression showed a consistent association between PCOS and hyperuricemia (MR = 5.23, = 1.4-26.26), obesity (MR = 3.26, CI = 1.33-8.01) and hypercholesterolemia (RM = 3.04, CI = 1.02-9.08). CONCLUSIONS: The study showed a higher prevalence of obesity, hypercholesterolemia, hypertriglyceridemia, hyperuricemia and diastolic hypertension in patients with PCOS than in the controls. No such thing for hyperglycaemia and systolic hypertension. Nevertheless, there are significant differences between both groups related to the BMI, WHI, systolic hypertension, hypercholesterolemia and hyperuricemia. A consistent association among the PCOS and hypercholesterolemia, obesity and hyperuricemia was found.

Epitheliocystis agents in sea bream Sparus aurata: morphological evidence for two distinct chlamydia-like developmental cycles.

The morphology of membrane-bound intracellular inclusions, or 'cysts', of epitheliocystis from sea bream Sparus aurata is described. Inclusions under the light microscope appear either granular or amorphous. Granular inclusions do not elicit a proliferative host reaction and contain the 3 distinctive developmental stages of chlamydial organisms: the highly pleomorphic reproductive form or reticulate body, the condensing form or intermediate body and the infective non-dividing rather uniform elementary body. Amorphous inclusions may elicit a proliferative host reaction and contain prokaryotic organisms which differ morphologically from those reported within granular cysts. More or less elongated electron-lucent organisms divide by fission to give rise to electron-dense non-dividing small cells with a dense nucleoid. Vacuolated and non-vacuolated small cells are reported. The morphology and developmental cycle of sea bream epitheliocystis agents would support their chlamydial nature; however, the immunohistochemical study conducted on gill samples which carried both inclusions failed to demonstrate the expression of lipopolysaccharide (LPS) chlamydial antigen. The different stages of the 2 distinct developmental cycles described in the present study are compared with electron microscope observations of epitheliocystis organisms reported from different host species. The hypothesis that epitheliocystis infection in the sea bream might be caused by a unique highly pleomorphic chlamydia-like agent, the life history of which includes 2 entirely different developmental cycles, is discussed.

[Hydatidiform mole. Clinical aspects, incidence and risk factors]. / Mola hidatiforme. Aspectos clínicos, incidencia y factores de riesgo.

From September 1st, 1992, to August 31st, 1994 35 patients with molar pregnancy were studied comparing them with 70 patients with normal pregnancy attended in the Hospital General de Zona 1 IMSS San Luis Potosí with the purpose of establishing the incidence of molar pregnancy to know the risk factors and to study the clinical aspects of it. The found incidence was: 1 molar pregnancy for each 415 deliveries, 22.85% of the mole patients were in a low socioeconomic position against 8.57% of the control group, suggesting that such condition is a risk factor. The average age was 26.28 years old in the mole patients and 26.42 years old in the control group. There were no differences in the number of first-pregnancy in both groups: 17.14% against 18.57% in the control group. The predominant symptoms and signs were: bleeding, vesicle expulsion, nausea and vomit. In all of them, a uterine curettage was performed, four needed blood transfusion, one needed subsequent curettage, there were no deaths. It is concluded that the incidence is similar to reports of other national authors, the low socioeconomic level is a factor for the aparition of this disease and it requires a precocious diagnostic to avoid complications.

[Post-salpingoclasia tubal recanalization. Preliminary report]. / Recanalización tubaria postsalpingoclasia. Informe preliminar.

PIP: 40 cases of microsurgery to reverse tubal sterilizations in a Mexican Social Security Institute (IMSS) hospital between March 1981-July 1984 were retrospectively studied to determine pregnancy rates and success factors. Only patients under 40 years old with no other causes of infertility were included. The same surgical team performed all the operations. 1-3 years of follow-up were provided. Patients' ages ranged from 20-40 years. 30 patients were 25-34 years old. 23 requested the reversal because of a 2nd marriage, 10 just to have another child, and 6 because of the death of 1 or more children. 31 of the sterilizations were by the Pomeroy method, 4 by fimbriectomy, 3 by Pomeroy and fimbriectomy, and 2 by Yoon rings. There were 10 pregnancies, 8 of which began in the 1st 6 postoperative months. 1 or both tubes were 5 cm or longer in all cases in which the woman became pregnant, and all of the sterilization were by the Pomeroy method. Radiographic control at about 6 months indicated that there was bilateral permeability in 9 other cases unilateral permeability in 8. Counting the cases of pregnancy, permeability was reestablished in 67.5% of cases. 2 pregnancies occurred in patients with isthmo-isthmic anastomoses and 8 in patients with isthmic-ampullar anastomoses. Microsurgery has been used in this service since 1981. It is anticipated that result will improve as the team becomes more experienced.^ieng