Post-immunobiotics increase resistance to primary respiratory syncytial virus infection and secondary pneumococcal pneumonia.

Previously, we demonstrated that post-immunobiotics derived from Lactobacillus gasseri TMT36, TMT39, and TMT40 strains (HK36, HK39 and HK40, respectively) differentially regulated Toll-like receptor 3 (TLR3)-mediated antiviral respiratory immunity in infant mice. In this work, we investigated whether the HK36, HK39 and HK40 nasal treatments were able to improve the resistance against primary respiratory syncytial virus (RSV) infection and secondary pneumococcal pneumonia. Our results demonstrated that the three treatments increased the resistance to primary viral infection by reducing variations in body weight, RSV titers and lung damage of infected infant mice. Post-immunobiotics significantly enhanced the expressions of interferon (IFN)-λ, IFN-ß, IFN-γ, interleukin(IL) - 1ß, IL-6, IL-27, Mx1, RNAseL and 2'-5'-oligoadenylate synthetase 1 (OAS1) genes and decreased tumour necrosis factor (TNF)-α in alveolar macrophages of RSV-challenged mice. In addition, the studies in the model of RSV-Streptococcus pneumoniae superinfection showed that the HK39 and HK40 treatments were capable of reducing lung damage, lung bacterial cell counts, and the dissemination of S. pneumoniae into the blood of infant mice. The protective effect was associated with increases in IFN-ß, IFN-γ, IL-10, and IL-27 in the respiratory tract. This study demonstrates that the nasal application of the post-immunobiotics HK39 and HK40 stimulates innate respiratory immunity and enhances the defences against primary RSV infection and secondary pneumococcal pneumonia offering an alternative to combat respiratory superinfections in children, which can be fatal.

New and rapid strategies for the diagnosis of bovine paratuberculosis "in situ" using latex particles.

The chemical coupling of a protoplasmatic antigen from Mycobacterium avium subsp. paratubeculosis onto core-shell carboxylated particles was investigated with the aim of producing latex-protein complexes to be used in immunoagglutination assays capable of detecting bovine paratuberculosis disease. For this purpose, sensitizations were carried out using both colored and not colored carboxylated latexes as well as the protoplasmatic antigen at pH close to its isoelectric point to favor the antigenic protein to approach the particle surface. In all cases, higher fractions of proteins were chemically-bound to carboxyl groups on the surface of the particles. The assessment of the performance of the visual immunoagglutination assays consisted of evaluating 111 sera from healthy and infected bovines with Mycobacterium avium subsp. paratuberculosis. Complexes obtained from the colored latex allowed an acceptable visual discrimination between the studied positive and negative sera. Most of the positive samples showed strong to very strong agglutination and only a few samples reacted weakly, i.e. a sensitivity of 70%. The specificity of the assay, on the other hand, was 86%. Therefore, this rapid detection technique allows an easy and inexpensive identification of animals possibly infected with paratuberculosis "in situ" in the herds.

Development and diagnostic validation of an ELISA based on an antigenic mixture for the detection of bovine tuberculosis.

Programs for the eradication of bovine tuberculosis (bTB) focus on the tuberculin skin test (TST) and slaughter of reactor cattle. However, the disease remains an animal health concern in several countries and improving the efficiency of the TST has become a critical issue. The detection of Mycobacterium bovis antibodies in serum, within weeks after the TST, may be a rapid and inexpensive way to improve bTB control. This study reports the validation of an enzyme-linked immunosorbent assay (ELISA) to detect bovine tuberculosis as an ancillary test to TST in dairy farms in Argentina. The estimated validation parameters were within the established requirements of the World Organization for Animal Health (OIE). The test demonstrated high repeatability, with coefficients of variation <25%. High test reproducibility through interlaboratory testing was also found, with an estimated Pearson coefficient of 0.9648 (95% confidence intervals 0.9315-0.9820). The ELISA detected tuberculous cattle unidentified by the TST. Of 43 animals sent to slaughterhouses that were ELISA positive 15-17 days after a negative TST, 36 were confirmed as infected with M. bovis by histopathology and IS6110 PCR. According to ROC curve analysis of results of 145 cattle from M. bovis-free herds and the 36 M. bovis-infected cattle, at a corrected optical density cut-off point of 0.3853, specificity was 95.95% and the positive predictive value at this cut-off was 83.72%. The ELISA detection test validated in this study could be readily applied in dairy farms, to complement a prior TST and improve livestock health.

Genetic diversity of Mycobacterium avium complex strains isolated in Argentina by MIRU-VNTR.

Mycobacterium avium sp. avium (MAA), M. avium sp. hominissuis (MAH), and M. avium sp. paratuberculosis (MAP) are the main members of the M. avium complex (MAC) causing diseases in several hosts. The aim of this study was to describe the genetic diversity of MAC isolated from different hosts. Twenty-six MAH and 61 MAP isolates were recovered from humans and cattle, respectively. GenoType CM® and IS1311-PCR were used to identify Mycobacterium species. The IS901-PCR was used to differentiate between MAH and MAA, while IS900-PCR was used to identify MAP. Genotyping was performed using a mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) scheme (loci: 292, X3, 25, 47, 3, 7, 10, 32) and patterns (INMV) were assigned according to the MAC-INMV database (http://mac-inmv.tours.inra.fr/). Twenty-two (22/26, 84·6%) MAH isolates were genotyped and 16 were grouped into the following, INMV 92, INMV 121, INMV 97, INMV 103, INMV 50, and INMV 40. The loci X3 and 25 showed the largest diversity (D: 0·5844), and the global discriminatory index (Hunter and Gaston discriminatory index, HGDI) was 0·9300. MAP (100%) isolates were grouped into INMV 1, INMV 2, INMV 11, INMV 8, and INMV 5. The HGDI was 0·6984 and loci 292 and 7 had the largest D (0·6980 and 0·5050). MAH presented a higher D when compared with MAP. The MIRU-VNTR was a useful tool to describe the genetic diversity of both MAH and MAP as well as to identify six new MAH patterns that were conveniently reported to the MAC-INMV database. It was also demonstrated that, in the geographical region studied, human MAC cases were produced by MAH as there was no MAA found among the human clinical samples.

Tumor inflamatorio de Pott en el siglo XXI: presentación de dos casos / Inflammatory Tumor of Pott in the 21st Century: Presentation of two cases

La sinusitis es una patología frecuente en la edad pediátrica y generalmente se presenta como complicación de una infección de vías respiratorias altas. En la era preantibiotica las sinusitis producían con cierta frecuencia, complicaciones extra o intracraneales. El tumor inflamatorio de Pott es una complicación, actualmente inusual de la sinusitis frontal entre sus causas más frecuentes. Sin embargo, el diagnóstico precoz y el tratamiento médico quirúrgico oportuno son fundamentales para evitar secuelas neurológicas devastadoras o incluso la muerte de los pacientes que padecen esta patología. Se presenta el caso de dos pacientes con diagnóstico de sinusitis, quienes desarrollan el tumor inflamatorio de Pott y sus complicaciones supurativas intracraneales extraaxiales. Se revisa en la literatura, las características clínicas, imagenologicas, diagnóstico y tratamiento de esta entidad.

Efecto de la Desnutrición Oculta Prenatal Sobre la Histología del Esplenio Callosal / Effect of Mild Protein Prenatal Malnutrition in the Callosal Splenium Histology

Ratas malnutridas prenatalmente con una dieta isocalórica y baja en proteínas, presentaron un menor diámetro axonal promedio en el esplenio callosal que los animales control, tanto de las fibras mielínicas como amielínicas. También se observó una mayor densidad axonal promedio, con respecto a los controles. Estas observaciones sugieren que: 1) las conexiones cortico-corticales (interhemisféricas) son vulnerables a la malnutrición proteica; y 2) lo anterior tendría incidencia en la velocidad de conducción interhemisférica, en particular con lo que dice relación con las conexiones visuales.

Adult rats malnourished prenatally with a low-protein, isocaloric diet showed smaller median fiber diameter of myelinated and unmyelinated fibers and a higher axonal density in the callosal splenium than controls. These findings suggest (i) that cortico-cortical (interhemispheric) connections are vulnerable to protein malnutrition; and (ii) this may affect interhemispheric conduction velocity, particularly in visual connections.

La malnutrición prenatal proteica leve afecta el desarrollo del cuerpo callaso anterior / Mild protein prenatal malnutrition specifically affects development of the anterior corpus callosum

Ratas mal nutridas prenatalmente y durante la lactancia con una dieta isocalórica y baja en proteínas, presentaron un cuerpo calloso más pequeño que los controles, a los 45-52 días de edad, lo cual fue consistente con las diferencias en los pesos cerebrales. En cambio, ratas mal nutridas prenatalmente y rehabilitadas durante la lactancia con una dieta alta en proteínas, mostraron normalidad en el peso cerebral y en el desarrollo de los tercios medio y posterior del cuerpo calloso. Estas observaciones sugieren: 1) que las conexiones cortico-corticales (interhemisféricas) son vulnerables a la malnutrición proteica; y 2) el cuerpo calloso anterior, que conecta áreas frontales a través de la línea media, es particularmente afectado por este tipo de malnutrición, a pesar de una rehabilitación dietaria durante la vida postnatal.

Pharmacokinetics and residues in milk of oxytetracyclines administered parenterally to dairy goats.

OBJECTIVE: To determine for two commercial preparations of oxytetracycline (OTC) the pharmacokinetic behaviour, the presence of detectable milk residues and the penetration in milk of OTC administered by intravenous (IV) (conventional formulation [CF]) and intramuscular (IM) routes (CF and long-acting [LA] formulations) in goats producing milk. The effects of these formulations on plasma activity values of creatine kinase (CK) and lactate dehydrogenase (LDH) were also determined as indicators of tissue damage. PROCEDURE: Five healthy lactating goats producing 1.5+/-0.5 L/d milk and weighing 56.0+/-4.8 kg were used. Single doses of OTC chlorhydrate (CF) were administered (20 mg OTC/kg) by IV (Trial 1 IV) and IM (Trial 1 IM) routes and OTC dehydrate (LA) by the IM route. The same goats were first given IV CF, then IM CF followed by IM LA with 3 weeks between each treatment. Blood and milk samples were taken. The quantification of OTC was performed by HPLC and the plasma activities of CK and LDH enzymes were determined by spectrophotometry. The presence of OTC residues in milk was determined by a commercial reagent. The plasma pharmacokinetic parameters were calculated using a two-compartment model. RESULTS: Estimates of kinetic variables following IV administration were: Vss= 400.0+/-120.0 mL/kg and CL= 110.0+/-14.0 (mL/h)/kg. The t(fi) for IV= 3.0+/-0.3 h; IM, CF = 10.5+/-2.1 h and IM, LA = 15.1+/-3.1 h. The concentration of OTC in milk at 48 h was: IV= 0.6+/-0.4; IM CF= 1.1+/-0.2 and at 72 h (IM LA)= 0.6+/-0.1 microg/mL and the penetration in milk of OTC was: IV= 70.0+/-18.0; IM CF= 79.0+/-14.0 and IM LA= 66.0+/-6.0%. The areas under the curve of CK and LDH activities in plasma were calculated by the trapezoidal method. Values of CK and LDH IM, LA were greater (P < 0.05) than those observed for IM, CF at 2 and 3 days after administration of the antibiotic. Finally, the bioavailability of OTC CF = 92.0+/-22.0 and LA= 78.0+/-23.0% was suitable for its usage by the IM route in lactating goats. CONCLUSION: Plasma concentration-time values of OTC administered parenterally in production dairy goats showed similar bioavailability for the two pharmaceutical preaprations. The presence of detectable residues in milk indicates that milk should not be used for human consumption for 2 and 3 days after administration of conventional and long-acting formulations, respectively. The increments in CK and LDH activities after the IM administration of LA are consistent with the presence of tissue damage provoked by the pharmaceutical preparations at the injection site.

Lesion of the bulbospinal noradrenergic pathways blocks desipramine-induced inhibition of the C-fiber evoked nociceptive reflex in rats.

Desipramine-induced inhibition of spinal cord nociceptive transmission was studied in rats with or without lesion of the bulbospinal noradrenergic system by recording the C-fiber evoked nociceptive reflex from a hind limb. Bulbospinal noradrenergic projections were lesioned by injecting intrathecally 20 microg of 6-hydroxydopamine 2 weeks before the electrophysiological experiments. Results show that desipramine (5, 10 and 20 mg/kg intraperitoneally) produced dose-dependent inhibition of the C reflex response duration in rats having intact noradrenergic bulbospinal systems. The inhibitory effect of desipramine was reduced or even abolished in rats pre-treated with 6-hydroxydopamine. In addition, [3H]-noradrenaline uptake was significantly lower in spinal cord slices arising from 6-hydroxydopamine lesioned animals, as compared to that from intact rats. These observations support the notion that the antinociceptive activity of antidepressants with noradrenergic selectivity depends on a normal rate of endogenous noradrenaline released by bulbospinal neurons.

Effects of mild protein prenatal malnutrition and subsequent postnatal nutritional rehabilitation on noradrenaline release and neuronal density in the rat occipital cortex.

There is evidence that severe malnutrition started during gestation and continued through lactation affects adversely the morphologic development of the neocortex leading to increased neuronal cell packing density and decreased dendritic branching. Nevertheless, the effect of purely mild protein prenatal malnutrition on neocortical development remains rather unexplored. This study evaluates the effects of mild protein prenatal malnutrition (8% casein diet, calorically compensated by carbohydrates) and subsequent postnatal nutritional rehabilitation (25% casein diet) on: (i) the potassium-induced release of [(3)H]-noradrenaline (NA) in occipital cortex slices obtained from 1- and 22-day-old pups; and (ii) the packing density of neurons in lateral, dorso-lateral and dorsal regions of the occipital cortex of 22-day-old pups by using the optical dissector method. The experiments were performed in rats normally fed during gestation and lactation (G(+)L(+)), malnourished during gestation but rehabilitated during lactation (G(-)L(+)) and malnourished during gestation and lactation (G(-)L(-)). At day 1 of age, no significant differences in body and brain weights were observed between prenatally well-nourished and malnourished pups. Nevertheless, at this early age, pups born from mothers submitted to the 8% casein diet had significantly higher cortical net percent NA release than pups born from mothers receiving the 25% casein diet. At weaning (22 days of age) G(-)L(+) rats had, compared to the G(+)L(+) group, similar body weight, brain weight and [(3)H]-NA release values, but significantly higher neuron density scores in the lateral region of the occipital cortex. In contrast, at 22 days of age, G(-)L(-) rats exhibited, compared to G(+)L(+) animals, significant deficits in body and brain weights as well as significant increases in cortical net percent NA release together with enhanced packing density of neurons in the lateral, dorso-lateral and dorsal regions of the occipital cortex. Moreover, in G(-)L(-) animals was not found the laterodorsal histogenetic gradient of neuronal cell packing density observed in G(+)L(+)rats. Results suggest that mild prenatal malnutrition per se is able to induce deleterious effects on cortical neuronal density, in spite of nutritional rehabilitation during lactation, through a mechanism involving central NA hyperactivity during gestation. Prosecution of malnutrition after birth magnifies both neurochemical and morphometric disorders.

Prenatal protein restriction alters synaptic mechanisms of callosal connections in the rat visual cortex.

Mild prenatal protein malnutrition, induced by reduction of the casein content of the maternal diet from 25 to 8%, calorically compensated by the addition of excess carbohydrates, leads to so-called "hidden" malnutrition in the rat. This form of malnutrition results in normal body and brain weights of pups at birth, but in significant alterations of their central nervous system neurochemical profiles. Since severe forms of prenatal malnutrition induce morpho-functional deficits on callosal interhemispheric communication together with brain neurochemical disturbances, we evaluated, in rats born from mothers submitted to an 8% casein diet, the potassium-induced release of [3H]-noradrenaline in visual cortex slices, as well as functional properties of callosal-cortical synapses by determining cerebral cortical excitability to callosal inputs and fatigability and temporal summation of transcallosal evoked responses. Rats born from mothers submitted to a 25% casein diet served as controls. At birth prenatally malnourished pups had significantly higher cortical percent net noradrenaline release (14.79 +/- 1.11) than controls (9.14 +/- 1.26). At 45-50 days of age, rehabilitated previously malnourished rats showed, when compared to controls; (i) significantly reduced percent net noradrenaline release in the visual cortex (4.50 +/- 0.52 vs 11.31 +/- 1.14); (ii) decreased cortical excitability to callosal inputs as revealed by significantly increased chronaxie (607.2 +/- 82.8 microseconds vs 351.3 +/- 47.7 microseconds); (iii) enhanced fatigability of transcallosal evoked responses as revealed by significantly decreased stimulus frequency required to fatigate the responses (4.9 +/- 0.8 Hz vs 9.2 +/- 1.3 Hz); and (iv) decreased ability of callosal-cortical synapses to perform temporal summation, as revealed by significantly reduced percent response increment to double-shock (54.2 +/- 6.2 vs 83.0 +/- 11.0, for a 3.2-ms interstimulus time interval). These changes, resulting from mild prenatal protein restriction, are discussed in relationship to developmental processes leading to the formation of synaptic contacts between callosal axons and their appropriate cortical target during perinatal age.

Biodegradabilidad aeróbica y anaeróbica del lixiviado de un relleno sanitario de Jujuy

Trabajo presentado en forma de afiche al 10o. Congreso Argentino de Saneamiento y Medio Ambiente, que mereció el premio AIDIS Argentina instituido para el mejor trabajo de esa categoría

Biodegradabilidad aeróbica y anaeróbica del lixiviado de un relleno sanitario de Jujuy

Trabajo presentado en forma de afiche al 10o. Congreso Argentino de Saneamiento y Medio Ambiente, que mereció el premio AIDIS Argentina instituido para el mejor trabajo de esa categoría

Prenatal malnutrition-induced functional alterations in callosal connections and in interhemispheric asymmetry in rats are prevented by reduction of noradrenaline synthesis during gestation.

Prenatal malnutrition results in increased concentration and release of central noradrenaline, a neurotransmitter that is an important regulator of normal regressive events such as axonal pruning and synaptic elimination. This suggests that some of the functional disturbances in brain induced by prenatal malnutrition could be due at least in part to increased noradrenaline activity that may enhance regressive events during early stages of development. To test this hypothesis we studied whether chronic administration of alpha-methyl-p-tyrosine, an inhibitor of tyrosine hydroxylase, to rats during gestation might prevent long-term deleterious effects of prenatal malnutrition on functional properties of interhemispheric connections of the visual cortex, and on asymmetry of visual evoked responses. The experiments were conducted on normal and malnourished rats 45-50 d of age. Prenatal malnutrition was induced by restricting the food consumption of pregnant rats to 40%, from d 8 postconception to parturition. At birth, prenatally malnourished rats had significantly greater whole-brain noradrenaline concentration as well as significantly enhanced noradrenaline release in the visual cortex. At 45-50 d of age, the malnourished group had a significantly smaller cortical area, exhibiting transcallosal evoked responses; in addition, the amplitude of these responses was significantly smaller. Malnourished rats showed a significant reduction of the normal interhemispheric asymmetry of visual evoked responses. The addition of 0.3% alpha-methyl-p-tyrosine to the diet of malnourished pregnant rats during the last 2 wk of gestation prevented functional disorders induced in the offspring by prenatal malnutrition on interhemispheric connectivity of visual areas and on interhemispheric bioelectrical asymmetry, probably by reducing the elevated brain noradrenaline activity and thereby restoring the normal trophic role of this neurotransmitter.

Biodegradabilidad aeróbica y anaeróbica del lixiviado de un relleno sanitario de Jujuy

Tomando en cuenta que las características de los líquidos de lixiviado que se acumulan en los sitios de disposición final dependen de numerosas variables, este trabajo se ha realizado con ensayos de biodegradabilidad de lixiviados de un relleno sanitario de la Pcia. de Jujuy, como uno de los pasos previos al diseño del sistema de descontaminación más apropiado para estos desechos

Enhancement of central noradrenaline release during development alters the packing density of neurons in the rat occipital cortex.

The effect of chronic yohimbine treatment early in life on packing density of neurons was evaluated in the occipital cortex of young rats. Yohimbine administration to pups between days 5 and 16 of postnatal life (2.5 mg/kg/day i.p.) resulted at 45 days of age in significantly higher neuronal density in layers II-V of the occipital cortex, the effect being more marked in the dorsal region than in the dorsolateral and lateral ones. Results suggest a relationship between enhanced central noradrenaline activity and altered development of the neuropil in the occipital cortex.

Central noradrenergic hyperactivity early in life: a hypothesis on the origin of morpho-functional brain disorders induced by malnutrition.

Many studies have revealed that malnutrition, caused either by insufficient or unbalanced diet, during early stages of growth and development could result in a variety of morpho-functional brain disturbances, whose severity depends on the time of onset, duration and intensity of the nutritional injury. Nevertheless, little is known about the intimate mechanisms by which early malnutrition impairs brain structure and function. This article reviews evidence showing that (i) developmental malnutrition induces central noradrenergic hyperactivity, (ii) noradrenaline exerts a trophic role during brain development, and (iii) pharmacological reduction of central noradrenergic hyperactivity prevents malnutrition-induced functional brain disturbances.

Central noradrenergic hyperactivity early in life: a hypothesis on the origin of morpho-functional brain disorders induced by malnutrition

Many studies have revealed that malnutrition, caused either by insufficient or unbalanced diet, during early stages of growth and development could result in a variety of morpho-functional brain disturbances, whose severity depends on the time of onset, duration and intensity of the nutritional injury. Nevertheless, little is known about the intimate mechanisms by which early malnutrition impairs brain structure and function. This article reviews evidence showing that (i) developmental malnutrition induces central noradrenergic hyperactivity, (ii) noradrenaline exerts a trophic role during brain development, and (iii) pharmacological reduction of central noradrenergic hyperactivity prevents malnutrition-induced functional brain disturbances

Tratamiento de efluentes de la industria vitivinícola

Desarrolla un sistema de tratamiento para los efluentes de la industria vitivinícola. Se implemento un programa de muestreo con un relevamiento previo de las condiciones en que se producen los efluentes parciales de las distintas etapas del proceso constituyendo, al reunirse, el efluente final. Se consideraron tres épocas en el proceso de vinificación según las características, en especial el grado de contaminación, de los líquidos residuales: molienda, clarificaciones y filtraciones, resto del ciclo. Se estableció que el sistema empleado debia constar de dos etapas: un tratamiento fisicoquímico y un tratamiento biológico con dos alternativas, lodos activados y filtro biológico. Con el tratamiento fisicoquímico-filtro biológico se obtuvo un efluente depurado, limpio e inodoro

Tratamiento de efluentes de la industria vitivinícola

Desarrolla un sistema de tratamiento para los efluentes de la industria vitivinícola. Se implemento un programa de muestreo con un relevamiento previo de las condiciones en que se producen los efluentes parciales de las distintas etapas del proceso constituyendo, al reunirse, el efluente final. Se consideraron tres épocas en el proceso de vinificación según las características, en especial el grado de contaminación, de los líquidos residuales: molienda, clarificaciones y filtraciones, resto del ciclo. Se estableció que el sistema empleado debia constar de dos etapas: un tratamiento fisicoquímico y un tratamiento biológico con dos alternativas, lodos activados y filtro biológico. Con el tratamiento fisicoquímico-filtro biológico se obtuvo un efluente depurado, limpio e inodoro