Bovine Tuberculosis and Brucellosis in Cattle and African Buffalo in the Limpopo National Park, Mozambique.

Bovine tuberculosis (BTB) and brucellosis are prevalent in buffaloes of the Kruger National Park (KNP, South Africa). Both diseases were considered to have no or a very low prevalence in wildlife and livestock in and around the Limpopo National Park (LNP, Mozambique). The same applies for tuberculosis in Gonarezhou National Park (GNP, Zimbabwe), but just recently, BTB was detected in buffaloes in the GNP and fears arose that the disease might also spread to the LNP as a result of the partial removal of the fences between the three parks to form the Great Limpopo Transfrontier Park. To assess the status of both diseases in and around LNP, 62 buffaloes were tested for bovine tuberculosis (BTB) and bovine brucellosis. The percentage of positive BTB reactors in buffalo was 8.06% using BovidTB Stat-Pak® and 0% with BOVIGAM® IFN-γ test and IDEXX ELISA. The brucellosis seroprevalence in buffalo was found to be 17.72% and 27.42% using Rose Bengal Test (RBT) and ELISA, respectively. In addition, 2445 cattle in and around the LNP were examined for BTB using the single intradermal cervical comparative tuberculin test (SICCT), and an apparent prevalence of 0.98% was found with no significant difference inside (0.5%) and outside (1.3%) the park. This is the first published report on the presence of positive reactors to BTB and bovine brucellosis in buffalo and cattle in and outside the LNP. Monitoring the wildlife-livestock-human interface of zoonotic high-impact diseases such as BTB and brucellosis is of outmost importance for the successful implementation and management of any transfrontier park that aims to improve the livelihoods of the local communities.

Mortality and translocation assay to study the protective capacity of Bifidobacterium lactis INL1 against Salmonella Typhimurium infection in mice.

The mouse has been largely used for the study of the protective capacity of probiotics against intestinal infections caused by Salmonella. In this work we aimed at comparing the mortality and translocation assay for the study of the protective capacity of the human breast milk-derived strain Bifidobacterium animalis subsp. lactis INL1 on a model of gut infection by Salmonella enterica subsp. enterica serovar Typhimurium. Different doses of S. Typhimurium FUNED and B. animalis subsp. lactis INL 1 were administered to Balb/c mice in a mortality or a translocation assay. The survival of the control group in the mortality assay resulted to be variable along experiments, and then we preferred to use a translocation assay where the preventive administration of 109 cfu of bifidobacteria/mouse for 10 consecutive days significantly reduced the number of infected animals and the levels of translocation to liver and spleen, with enhanced secretory immunoglobulin A and interleukin 10 production in the small and large intestine, respectively. Ten days of B. animalis subsp. lactis strain INL1 administration to mice significantly reduced both the incidence and the severity of Salmonella infection in a mouse model of translocation. This work provided the first evidence that a translocation assay, compared to a mortality assay, could be more useful to study the protective capacity of probiotics against Salmonella infection, as more information can be obtained from mice and less suffering is conferred to animals due to the fact that the mortality assay is shorter than the latter. These facts are in line with the guidelines of animal research recently established by the National Centre for the Replacement, Refinement & Reduction of Animals in Research.

Cell viability and immunostimulating and protective capacities of Bifidobacterium longum 51A are differentially affected by technological variables in fermented milks.

AIM: To investigate the cell viability of Bifidobacterium longum 5(1A) in fermented milks and to study its immunostimulating and protective capacity against Salmonella enterica ssp. enterica serovar Typhimurium infection in mice. METHODS AND RESULTS: Bifidobacterium longum 5(1A) was added to milk fermented with different yoghurt starter cultures, before or after fermentation, and viability was monitored during storage (5°C, 28 days). Resistance to simulated gastric acid digestion was assessed. Fermented milks were orally administered to mice for 10 days followed by oral infection with Salmonella Typhimurium. The number of IgA+ cells in the small and large intestine was determined before infection. Survival to infection was monitored for 20 days. Bifidobacterium longum 5(1A) lost viability during storage, but the product containing it was effective for the induction of IgA+ cells proliferation in the gut and for the protection of mice against Salm. Typhimurium infection. CONCLUSIONS: Cell viability of Bif. longum 5(1A) in fermented milks along storage did not condition the capacity of the strain to enhance the number of IgA+ cells in the gut and to protect mice against Salmonella infection. SIGNIFICANCE AND IMPACT OF THE STUDY: The uncoupling of cell viability and functionality demonstrated that, in certain cases, nonviable cells can also exert positive effects.

Preservation of functionality of Bifidobacterium animalis subsp. lactis INL1 after incorporation of freeze-dried cells into different food matrices.

The aim of this work was to investigate how production and freeze-drying conditions of Bifidobacterium animalis subsp. lactis INL1, a probiotic strain isolated from breast milk, affected its survival and resistance to simulated gastric digestion during storage in food matrices. The determination of the resistance of bifidobacteria to simulated gastric digestion was useful for unveiling differences in cell sensitivity to varying conditions during biomass production, freeze-drying and incorporation of the strain into food products. These findings show that bifidobacteria can become sensitive to technological variables (biomass production, freeze-drying and the food matrix) without this fact being evidenced by plate counts.

Modificaciones estructurales en el estómago y el hígado de Paralabrax maculatofasciatus (Steindacher, 1868) ante situaciones de estrés crónico / Structural modifications in the stomach and liver of Paralabrax maculatofasciatus (Steindacher, 1868) under chronic stress conditions

Para determinar el efecto del estrés a nivel del aparato digestivo, organismos de la especie Paralabrax maculatofasciatus fueron sometidos a dos modelos de estrés: el primero por descenso en el nivel del agua (n=84) por 30 y 60 minutos diariamente, y el segundo por cultivo a altas densidades (n=96) de 1,6 y 3,2 kg/100 1. Se tomaron muestras de estómago e hígado y se fijaron en formol al 10%. Los tejidos fueron incluidos en parafina, realizándose cortes de 6 um que se tiñeron con H-E. En ambas condiciones de estrés el estómago presentó cambios morfológicos importantes tales como hipertrofia y atrofia en la capa mucosa. En el hígado se observaron cambios de coloración y textura, así como hemolisis, inflamación y necrosis. Por descenso en el nivel del agua el estómago presentó diferencias morfométricas significativas (P<0,05), entre los diferentes tiempos de exposición al estrés y/o entre los días de muestreo en algunos de los parámetros medidos. Por cultivo a altas densidades, se observaron diferencias significativas en algunos parámetros, 4 de ellos presentaron interacción densidad-muestreo y el resto presentaron diferencias significativas sin interacción. El hígado presentó diferencias significativas (P<0,05) en el perímetro nuclear entre los diferentes días de muestreo. El estrés provocado por alta densidad de cultivo resultó ser el factor que provocó daños tisulares más severos en los órganos estudiados, de ahí la importancia de mantener las condiciones adecuadas en los cultivos, como son el nivel de agua y el número de organismos por estanque.

To determine stress effects at histological level in the digestive system, organisms of Paralabrax maculatofasciatus were submitted to 2 stress models. One of decreasing the water level (n=84) for 30 and 60 min daily, and another at high density cultures (n=96) of 1.6 and 3.2 Kg/1001. Stomach and the liver samples were fixed in 10% formalin, embedded in paraffin wax, sectioned at 6 urn and stained with haematoxylin-eosin. In both stress conditions, the stomach showed important morphological changes in the mucosa layer, such as hypertrophy and atrophy. In the liver, coloration and texture changes were observed as well as haemolisis, inflammation and necrosis. In the water decrease model, the mucosa layer of stomach showed significant morphometric differences (P<0.05), between different times of stress exposure and/or between different days, in some of the measured parameters. In the high-density model, significant differences in the stomach mucosa showed a sampling-density interaction in 4 parameters and 6 other parameters showed differences with no interaction. Liver showed significant differences in nuclear perimeter between sampling-days. Stress caused by high-density culture proved to be the factor that caused the most serious tissue damage.

17ß-Oestradiol indirectly inhibits thyrotrophin-releasing hormone expression in the hypothalamic paraventricular nucleus of female rats and blunts thyroid axis response to cold exposure.

Energy expenditure and thermogenesis are regultated by thyroid and sex hormones. Several parameters of hypothalamic-pituitary-thyroid (HPT) axis function are modulated by 17ß-oestradiol (E(2)) but its effects on thyrotrophin-releasing hormone (TRH) mRNA levels remain unknown. We evaluated, by in situ hybridisation and Northern bloting, TRH expression in the paraventricular nucleus of the hypothalamus (PVN) of cycling rats, 2 weeks-ovariectomised (OVX) and OVX animals injected s.c. during 1-4 days with E(2) (5, 50, 100 or 200 µg / kg) (OVX-E). Serum levels of E(2), thyroid-stimulating hormone (TSH), prolactin, corticosterone and triiodothyronine (T(3)) were quantified by radioimmunoassay. Increased serum E(2) levels were observed after 4 days injection of 50 µg / kg E(2) (to 68.5 ± 4.8 pg / ml) in OVX rats. PVN-TRH mRNA levels were slightly higher in OVX than in virgin females at dioestrous 1 or pro-oestrous, decreasing proportionally to increased serum E(2) levels. E(2) injections augmented serum T(3), prolactin, and corticosterone levels. Serum TSH levels augmented with 4 days 50 µg / kg E(2), but not with the higher doses that enhanced serum T(3) levels. Exposure to cold for 1 h resulted in marked HPT axis activation in OVX rats, increasing the levels of TRH mRNA along the rostro-caudal PVN areas, as well as serum TSH, T(3), corticosterone and prolactin levels. By contrast, no significant changes in any of these parameters were observed in cold-exposed OVX-E (50 µg / kg E(2)) rats. Very few PVN-TRHergic neurones expressed the oestrogen receptor type-α, suggesting that the effects of E(2) on PVN-TRH expression are indirect, most probably as a result of its multiple modulatory effects on circulating hormones and their receptor sensitivity. The blunted response of OVX-E rats to cold coincides with the effects of E(2) on the autonomic nervous system and increased cold tolerance.

Differential responses of thyrotropin-releasing hormone (TRH) neurons to cold exposure or suckling indicate functional heterogeneity of the TRH system in the paraventricular nucleus of the rat hypothalamus.

Thyrotropin-releasing hormone (TRH) is released from the median eminence upon neural stimulation such as cold or suckling exposure. Concomitant with the cold- or suckling-induced release of TRH is a rapid and transient increase in the expression of proTRH mRNA in the paraventricular nucleus (PVN) of the hypothalamus. We employed two strategies to determine whether TRH neurons responding to cold exposure are different from those responding to suckling. First, we attempted to identify a marker of cellular activation in TRH neurons of the PVN. Cold induced c-fos expression in about 25% of TRH neurons of the PVN, but no induction was observed by suckling. Moreover, we explored the expression of a variety of immediate early genes including NGFI-A, fra-1 and c-jun, or CREB phosphorylation but found none to be induced by suckling. The number of cells expressing high levels of proTRH mRNA was counted and compared to total expressing cells. An increased number of cells expressing high levels of proTRH mRNA was observed when both stimuli were applied to the same animal, suggesting that different cells respond separately to each stimulus. We therefore analyzed the distribution of responsive TRH neurons as defined by the cellular level of proTRH mRNA. The proTRH mRNA signal was analyzed within three rostrocaudal zones of the PVN and within six mediolateral columns. Results showed that in response to cold, all areas of the PVN of the lactating rat present increased proTRH mRNA levels, including the anterior zone where few hypophysiotropic TRHergic cells are believed to reside. The distribution of the proTRH mRNA expressing cells in response to cold was quite comparable in female and in male rats. In contrast, the response after suckling was confined to the middle and caudal zones. Our results provide evidence of a functional specialization of TRH cells in the PVN.

Optical alignment of segmented mirrors to the fluorescence detectors of the Pierre Auger observatory.

Segmented mirrors will be used in the telescopes of the Pierre Auger Fluorescence detector. To align the segments, we have developed four methods in which (a) the image of the stop border, (b) the image of a screen with concentric circles, and (c) the Ronchi pattern are used. In addition to these, we have developed a new method, (d), in which instead of the Ronchi ruling, we have used a circular grid. In this case we obtain a moiré pattern for each segment by means of which the experimental setup is simplified, and the sensitivity of the alignment is improved.

n-Hindle-sphere arrangement with an exact ray trace for testing hyperboloid convex mirrors.

We present calculations with an exact ray trace to determine the dimensions that define one or two Hindle spheres, since the paraxial theory is incongruent for convex hyperboloid mirrors with small f numbers. The equations are generalized to calculate the dimensions of n Hindle spheres, since in this way it is possible to reduce the dimensions of the spheres more. Actual calculations are done for the secondary mirrors of the Benemerita Universidad Autonoma de Puebla and Large Milimetric Telescopes; experimental results are shown for the latter.

Ronchi test with a square grid.

We use a square grid in the Ronchi test. This grid allows processing of both the X and the Y directions when calculating optical path difference. We use trapezoidal integration to analyze the new patterns, since it does not have the smoothing drawback at the edges of the wave front.