GABA-ergic control of prolactin release in rainbow trout (Oncorhynchus mykiss) pituitaries in vitro

The involvement of γ-aminobutyric acid (GABA) in the control of prolactin (PRL) release was investigated in rainbow trout using both perifused pituitary fragments and pituitary cells in primary culture. In our perifusion system, infusion of GABA (10⁻⁶ to 10⁻⁴ M) caused an inhibition of PRL release (between 20 and 40%). Administration on perifused pituitary fragments of 3APS, a GABAa agonist, mimicked this inhibitory effect. Moreover, bicuculline, a specific antagonist of GABAa receptors, totally abolished GABA effect. When tested on cultured pituitary cells during 40h exposure, GABA (10⁻⁵ M) caused a significant decrease in PRL release (24.5%). Baclofen, a specific agonist for GABAb receptor tested at 10⁻⁶ and 10⁻⁵ M, also inhibited PRL released from cultured pituitary cells. These results demonstrate that GABA inhibits PRL release by acting directly on pituitary cells and that probably both types of GABA receptor (a and b) are involved in this regulation.

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Résumé Nous avons étudié l'implication de l'acide γ-aminobutirique (GABA) dans le controle de la sécrétion de prolactine (PRL) chez la truite arc-en-ciel en utilisant à la fois des fragments d'hypophyse perifusés et des cellules hypophysaires en culture. Dans notre système de périfusion, le GABA (10⁻⁶ à 10⁻⁴ M) inhibe la libération de PRL (entre 20 et 40%). L'administration sur les fragments d'hypophyse périfusés de 3APS, un agoniste des récepteurs GABAa, reproduit ces effets inhibiteurs. De plus, la bicuculline, un antagoniste spécifique des récepteurs de type GABAa, abolie complètement les effets du GABA. Lorsqu'il est testé pendant 40h sur des cellules en culture, le GABA (10⁻⁵ M) réduit de manière significative la libération de PRL (24.5%). Le Baclofen, un agoniste spécifique des récepteurs GABAb testé à 10⁻⁶ et 10⁻⁵ M, inhibe aussi la libération de PRL par les cellules en culture. Ces résultats démontrent que le GABA inhibe la libération de PRL en agissant directement sur les cellules hypophysaires et que les 2 types de récepteurs GABA (a et b) sont impliqués dans cette régulation.

     

Inhibition of apple polyphenol oxidase activity by procyanidins and polyphenol oxidation products

The rate of consumption of dissolved oxygen by apple polyphenol oxidase in cider apple juices did not correlate with polyphenol oxidase activity in the fruits and decreased faster than could be explained by the decrease of its polyphenolic substrates. The kinetics parameters of a crude polyphenol oxidase extract, prepared from apple (Braeburn cultivar), were determined using caffeoylquinic acid as a substrate. Three apple procyanidin fractions of n 80, 10.5, and 4 were purified from the parenchyma of cider apples of various cultivars. Procyanidins, caffeoylquinic acid, (-)-epicatechin, and a mixture of caffeoylquinic acid and (-)-epicatechin were oxidized by reaction with caffeoylquinic acid o-quinone in order to form oxidation products. All the fractions were evaluated for their inhibitory effect on PPO activity. Native procyanidins inhibited polyphenol oxidase activity, the inhibition intensity increasing with n. The polyphenol oxidase activity decreased by 50% for 0.026 g/L of the fraction of n 80, 0.17 g/L of the fraction of n 10.5, and 1 g/L of the fraction of n 4. The inhibitory effect of oxidized procyanidins was twice that of native procyanidins. Oxidation products of caffeoylquinic acid and (-)-epicatechin also inhibited polyphenol oxidase.     

Assessing the fertility of two mares cloned from the same founder animal

Two cloned mares, produced from the same sample of skin fibroblasts, were bred during four breeding seasons from their second year of age, as embryo donors, in exactly the same conditions, using the same stallions for both cloned mares. The aim of this study was to test the embryo donor potential of cloned mares and to compare the results obtained from two cloned mares of the same mare with other embryo donor mares (n = 31–39 per breeding season) at the same stud. For both cloned mares, 19 embryos were recovered by 43 collection attempts (44%) (7/22 for one; 12/21 for the other), 16 (84%) pregnancies (5/7 for one, 11/12 for the other) were obtained at day 14 post-ovulation (D₁₄), and 12 (3/7 for one; 9/12 for the other) foals were born. One cloned mare was a less efficient donor mare than the other (p < .05), In control donor mares, 623 embryo collections were performed, with a recovery rate (80%—496/623) significantly higher than for cloned mares. The recovery rate in the subpopulation of 2–5-year-old control donor mares (same age of cloned mares) (89%—127/143) and The recovery rate in the subpopulation of 12 control mares bred with the seven same stallions as clones (55%—17/31), were both higher than for cloned mare (p < .05). The success rate of transfer was not different between embryos produced by cloned mares (84%—16/19) and those produced by control donor mares (79%—392/496). However, the foaling rate per embryo collection was significantly lower for cloned mares (28%—12/43) than for control donor mares (52% - 325/623) (p < .05).     

Fagacées: a tree-centered growth and yield model for sessile oak (<Emphasis Type="Italic">Quercus petraea</Emphasis> L.) and common beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.)

• Introduction  

Fagacées is a growth model that has been developed for pure Oak or Beech stand management in even-aged high forests and coppice with standards forests. It has been calibrated for the plain forests of northern France.     

Isolation and characterisation of a bacterial strain degrading the herbicide sulcotrione from an agricultural soil

BACKGROUND: The dissipation kinetics of the herbicide sulcotrione sprayed 4 times on a French soil was studied using a laboratory microcosm approach. An advanced cultivation‐based method was then used to isolate the bacteria responsible for biotransformation of sulcotrione. Chromatographic methods were employed as complementary tools to define its metabolic pathway. RESULTS: Soil microflora was able quickly to biotransform the herbicide (DT₅₀ ≈ 8 days). 2‐Chloro‐4‐mesylbenzoic acid, one of its main metabolites, was clearly detected. However, no accelerated biodegradation process was observed. Eight pure sulcotrione‐resistant strains were isolated, but only one (1OP) was capable of degrading this herbicide with a relatively high efficiency and to use it as a sole source of carbon and energy. In parallel, another sulcotrione‐resistant strain (1TRANS) was shown to be incapable of degrading the herbicide. Amplified ribosomal restriction analysis (ARDRA) and repetitive extragenic palendromic PCR genomic (REP‐PCR) fingerprinting of strains 1OP and 1TRANS gave indistinguishable profiles. CONCLUSION: Sequencing and aligning analysis of 16S rDNA genes of each pure strain revealed identical sequences and a close phylogenetic relationship (99% sequence identity) to Pseudomonas putida. Such physiological and genetic properties of 1OP to metabolise sulcotrione were probably governed by mobile genetic elements in the genome of the bacteria. Copyright © 2011 Society of Chemical Industry     

Detection of Coxiella burnetii, the agent of Q fever, in oviducts and uterine flushing media and in genital tract tissues of the non pregnant goat

The aim of the present study was the detection and quantification of Coxiella burnetii DNA in the flushing media (oviducts and uterine horns) and genital tract tissues of non pregnant goats from 20 goats chosen at random from 86 goats originating from 56 different breeding herds in south-west France. The serological prevalence rate of C. burnetii in the study population was 70.3%.The DNA of C. burnetii was identified using conventional PCR in the flushing media from the oviducts and uterus in 8/20 goats (40%) and in genital tract tissues (oviduct, uterus and ovary) in 5/20 goats (25%). This study clearly shows for the first time that the media used to flush the oviducts or uterine horns, collected using the standard embryo harvesting technique in goats, are susceptible to infection with C. burnetii. The 16 conventional PCR-positive samples were also analyzed using real-time PCR. The bacterial load of the oviduct and uterine flushing media varied from 2.9 × 10⁴ to 7.5 × 10⁶ bacteria per flushing medium, while the bacterial load of the tissue samples varied from 1.0 × 10² to 1.5 × 10⁵ bacteria per mg of tissue. The infection of genital tract flushing media and tissues is a risk factor for the transmission of C. burnetii from donor to recipient during embryo transfer or to the embryo and fetus when gestation is pursued to term.     

Genetic immunisation of cattle against bovine herpesvirus 1: glycoprotein gD confers higher protection than glycoprotein gC or tegument protein VP8

Bovine herpesvirus 1 (BoHV-1) has frequently been used as a model for testing parameters affecting DNA immunisation in large animals like cattle. However, the selection of target antigens has been poorly studied, and most of the experiments have been conducted in mice. In the present study, we demonstrated in cattle that a DNA vaccine encoding BoHV-1 glycoprotein gD induces higher neutralising antibody titres than vaccines encoding BoHV-1 gC. Additionally, we show that a DNA vaccine encoding a secreted form of gD induces a higher immune response than a vaccine encoding full-length gD. However, the enhanced immunogenicity associated with the secretion of gD could not be extended to the glycoprotein gC. The current study also describes for the first time the development and the evaluation of a DNA vaccine encoding the major tegument protein VP8. This construct, which is the first BoHV-1 plasmid vaccine candidate that is not directed against a surface glycoprotein, induced a high BoHV-1 specific cellular immunity but no humoral immune response. The calves vaccinated with the constructs encoding full-length and truncated gD showed a non-significant tenfold reduction of virus excretion after challenge. Those calves also excreted virus for significantly (p < 0.05) shorter periods (1.5 days) than the non-vaccinated controls. The other constructs encoding gC and VP8 antigens induced no virological protection as compared to controls. Altogether the DNA vaccines induced weaker immunity and protection than conventional marker vaccines tested previously, confirming the difficulty to develop efficient DNA vaccines in large species.     

Influence of the route of administration on efficacy and tissue distribution of ivermectin in goat

The tissue concentration and efficacy of ivermectin after per os and subcutaneous administration were compared in goats experimentally infected with Trichostrongylus colubriformis (ivermectin-susceptible strain, INRA). Infected goats (n = 24) were treated per os (n = 9) or subcutaneously (n = 9) with ivermectin, 0.2 mg/kg, or kept as not treated controls. The faecal egg counts and small intestine worm counts were determined. Ivermectin concentration was measured in the plasma, gastrointestinal tract, lung, skin or hair, liver and adipose tissues at 0, 2, 7 and 17 days post-treatment. The efficacy of ivermectin against T. colubriformis infection in goat was 98.7 and 99.9% for subcutaneous and oral administration, respectively. Ivermectin concentration declined with time and only residual concentration was measured at 17 days post-treatment in plasma and gastrointestinal tract. Ivermectin concentration was higher after subcutaneous compared to per os injection in most of the tissue examined. In skin, hair and subcutaneous adipose tissue ivermectin persisted at significant concentrations 17 days post-treatment for both routes of administration. In our experimental conditions, ivermectin provides similar efficacy against T. colubriformis after subcutaneous or per os administration in goat. However, the lower ivermectin levels in tissues after per os administration suggest that the lasting of efficacy may be shortened after per os compared to subcutaneous administration especially in animals with poor body condition in pasture where re-infection occurs quickly after anthelmintic treatment.     

Health hazards for terrestrial vertebrates from toxic cyanobacteria in surface water ecosystems

Toxigenic cyanobacteria are photosynthetic prokaryotes that are most often recognized in marine and freshwater systems, such as lakes, ponds, rivers, and estuaries. When environmental conditions (such as light, nutrients, water column stability, etc.) are suitable for their growth, cyanobacteria may proliferate and form toxic blooms in the upper, sunlit layers. The biology and ecology of cyanobacteria have been extensively studied throughout the world during the last two decades, but we still know little about the factors and processes involved in regulating toxin production for many cyanobacterial species. In this minireview, we discuss these microorganisms, and more especially the toxins they produce, as a potential and important health risk for wild and domestic animals.     

SLO angiography: arterio-venous filling times in monkey and minipig

Confocal scanning laser ophthalmosocope (cSLO) is a new technique which enables ocular fundus image recording and dynamic retinal angiography to be performed. The ocular fundus image is acquired sequentially, point by point, and is reconstructed on a video monitor at the rate of 25 images per second. The aim of this paper is to evaluate the feasibility of measuring retinal arterio‐venous filling times (AVFT) with a I + Tech cSLO. Three young adult cynomolgus monkeys and three young adult Göttingen minipigs were used as experimental models. All animals were anesthetized using a zolazepam + tiletamine mixture injected intramuscularly; heart rate and rectal temperature were monitored and corneal irrigation was regularly performed. For all subjects, prior to examination, hematocrit and globe axial length were measured. The images were recorded, stabilized and analyzed. The retinal examination consisted of retinal images with 40° field cSLO, retinal fluorescein angiography and arterio‐venous 50% filling time measurements. For each subject all images were easily recorded while keeping the animals in a normally lighted room without having to use any additional optical device. AVFT using an I + Tech cSLO is easily performed in monkeys and minipigs. AVFT measurements in minipigs and monkeys are similar. These results suggest that minipigs can replace monkeys as an experimental species for AVFT investigations.