Cultivation of monoxenous trypanosomatids: A minireview.

Trypanosomatids are obligatory parasites, some of which are responsible for important human and animal diseases, but the vast majority of trypanosomatids are restricted to invertebrate hosts. Isolation and in vitro cultivation of trypanosomatids from insect hosts enable their description, characterization, and subsequently genetic and genomic studies. However, exact nutritional requirements are still unknown for most trypanosomatids and thus very few defined media are available. This mini review provides information about the role of different ingredients, recommendations and advice on essential supplements and important physicochemical parameters of culture media with the aim of facilitating first attempts to cultivate insect-infesting trypanosomatids, with a focus on monoxenous trypanosomatids.

Identifying an Optimal Screen Mesh to Enable Augmentorium-Based Enhanced Biological Control of the Olive Fruit Fly Bactrocera oleae (Diptera: Tephritidae) and the Mediterranean Fruit Fly Ceratitis capitata (Diptera: Tephritidae).

The augmentorium is a cost-effective screened enclosure designed to receive fruits infested with insect pests, retain the pests inside but let their natural enemies escape to enhance biological control of pest populations. Screen selection is critical to ensure that an augmentorium is effective for a particular system. Here we tested five types of screens with a mini-augmentorium design and measured the escape of four insect species under laboratory conditions: the pests olive fruit fly Bactocera oleae (Rossi) (Diptera: Tephritidae) and Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), and the parasitoids Psyttalia lounsburyi (Silvestri) (Hymenoptera: Braconidae) and Psyttalia ponerophaga (Silvestri) (Hymenoptera: Braconidae). The sex ratio of insects that escaped the screens was compared to the sex ratio of insects that could not escape. Results showed that one screen type ('crystal mesh') was the best for the purpose of designing a functional augmentorium: it retained 90% of B. oleae adults and 100% of C. capitata adults while letting 72% of Psyttalia lounsburyi adults and 94% of P. ponerophaga adults escape. The other screen types tested were suboptimal, either because they let too many flies freely escape or because they retained too many parasitoids. Sex ratio was almost always similar for insects that managed to escape the screens and insects that were retained, except for P. ponerophaga and the screen type 'light mesh'. These results are promising for the development of a functional augmentorium against the olive fruit fly and further implementation against Mediterranean fruit fly.

First Report of Grapevine Pinot gris virus (GPGV) in grapevine in France.

Grapevine Pinot gris virus (GPGV), belonging to the genus Trichovirus of the family Betaflexiviridae, was first identified by siRNA sequencing in northern Italy in 2012, in the grapevine varieties Pinot gris, Traminer, and Pinot Noir, which exhibited mottling and leaf deformation (1), and in asymptomatic vines, with a lower frequency. Since 2012, this virus has also been reported in South Korea, Slovenia, Greece (3), Czech Republic (2), Slovakia (2), and southern Italy (4). In 2014, GPGV was identified by Illumina sequencing of total RNAs extracted from leaves of the Merlot variety (Vitis vinifera) grafted onto Gravesac rootstock originated from a vineyard in the Bordeaux region of France. This Merlot plant exhibited fanleaf-like degeneration symptoms associated with Tomato black ring virus (TBRV) infection. Cuttings were collected in 2010 and maintained thereafter in a greenhouse. The full-length genome was assembled either de novo or by mapping of the Illumina reads on a reference GPGV genome (GenBank FR877530) using the CLC Genomics workbench software (CLC Bio, Qiagen, USA). The French GPGV isolate "Mer" (7,223 nucleotides, GenBank KM491305) is closely related to other European GPGV sequences; it exhibits 95.4% nucleotide identity with the reference Italian isolate (NC_015782) and 98 to 98.3% identity with Slovak isolates (KF134123 to KF134125). The higher divergence between French and Italian GPGV isolates was mainly due to differences in the 5' extremity of the genome, as already shown with the Slovak GPGV isolates. RNA extracted from phloem scrapings of 19 cv. Merlot vines from the same plot collected in 2014 were analyzed by RT-PCR using the specific primer pair Pg-Mer-F1 (5'-GGAGTTGCCTTCGTTTACGA-3') and Pg-Mer-R1 (5'-GTACTTGATTCGCCTC GCTCA-3'), designed on the basis of alignments of all available GPGV sequences from GenBank. The resulting amplicon of 770 bp corresponded to a fragment of the putative movement protein (MP) gene. Seven (35%) of the tested plants gave a strong positive amplification. Three RT-PCR products were directly sequenced and showed 99.3 to 99.5% identity within the MP gene of the GPGV-Mer isolate. Given the mixed viral infection status of the vines found infected by GPGV, it was not possible to associate a specific symptomatology with the presence of GPGV. Furthermore, similar RT-PCR tests were also performed on RNA extracts prepared from two plants of cv. Carignan that originated from a French grapevine collection, exhibiting fanleaf-like symptoms without any nepovirus detection. These samples similarly gave a strong positive amplification. The sequences obtained from the two Carignan vines showed 98.4 and 97.8% identity with the GPGV-Mer isolate. To our knowledge, this is the first report of GPGV in France. GPGV has been discovered in white and red berry cultivars, suggesting that its prevalence could be important in European vineyards (2). Further large-scale studies will be essential to determine the world prevalence of GPGV and to evaluate its potential effects on yield and on wine quality, as well as to shed light on GPGV epidemiology. Of particular concern is whether, like the other grapevine-infecting Trichovirus, Grapevine berry inner necrosis virus (GPGV) can be transmitted by the eryophid mite Colomerus vitis. References: (1) A. Giampetruzzi et al. Virus Res. 163: 262, 2012. (2) M. Glasa et al. Arch. Virol. 159: 2103, 2014. (3) G. P. Martelli, J. Plant Pathol. 96: S105, 2014. (4) M. Morelli et al. J. Plant Pathol. 96:431, 2014.

First Report of Grapevine redglobe virus (GRGV) in Grapevine in France.

The isometric virus Grapevine redglobe virus (GRGV), was first described on grapevine cv. Red Globe in southern Italy in 2000 (3) and later in Greece and California. GRGV belongs to the genus Maculavirus in the family Tymoviridae. These viruses are thought to be disseminated through propagation and grafting, as no vectors or seed transmission are known to date. A partial sequence (2,006 nucleotides [nt]) encompassing the 3' end of the replicase, the coat protein, and P17 genes, was obtained in 2003 (1). GRGV infections are apparently symptomless (2). In 2014, GRGV was identified by Illumina sequencing of total RNAs extracted from a Vitis vinifera cv. Cabernet franc (CF) vine grafted onto Gravesac in a vineyard of the Bordeaux region in France. This Cabernet franc plant displayed fanleaf-like degeneration symptoms associated with Tomato black ring virus (TBRV) infection. It had been collected in 2010 and maintained since in a greenhouse. The partial contigs assembled from the Illumina reads (552 and 430 nt, both in the putative replicase gene, KM491303 and KM491304) showed 85.9 and 86.3% nt identity with the partial sequence of a GRGV Italian isolate (AF521577), respectively. Total RNA extracts from leaves of 18 plants of cv. Cabernet franc from the same plot, collected in 2014, were analyzed by RT-PCR using specific primers RG-CF-F1 (5'-GAATTCGCTGTCGGCCACTC-3') and RG-CF-R1 (5'-AGTGAGAGGAGAGATTCCATC-3') designed on the basis of the alignment of the partial sequences of GRGV-CF and the Italian isolate (AF521577). Fifteen (83%) of the plants gave strong positive amplification for GRGV. Given the mixed viral infection status of these vines, it was not possible to associate a specific symptomatology with the presence of GRGV. Two RT-PCR amplicons were directly sequenced and showed 91.5 and 91.7% identities, respectively, with the reference GRGV-CF sequence. To our knowledge, this is the first report of GRGV in France. Further studies will be necessary to determine the prevalence of GRGV in the French vineyards and varieties, including rootstocks, and its possible threat to the grapevine industry. Studies are also needed to assess the pathogenicity of GRGV. Similarly to its close relative, Grapevine fleck virus, does it induce latent or semi-latent infections in Vitis vinifera and rootstock hybrids, influencing vigor, rooting ability, and graft compatibility? References: (1) N. Abou Ghanem-Sabanadzovic et al. Virus Genes 27:11, 2003. (2) G. P. Martelli et al. Arch. Virol. 147:1847, 2002. (3) S. Sabanadzovic et al. Arch. Virol. 145:553, 2000.

Growth biostimulation of quorum-quenching bacteria by gammagamma-heptalactone treatment in the hydroponic rhizosphere of Solanum tuberosum.

Several bacterial plant pathogens, including Pectobacterium, use a cell-to-cell communication system called quorum sensing (QS) to synchronize and regulate expression of the virulence factors. In this study, the biomolecule gamma-heptalactone (GHL) was introduced in hydroponic culture of Solanum tuberosum to stimulate growth of the native rhizospheric bacteria which are able to degrade the QS signal, hence potentially quench the QS-regulated virulence of Pectobacterium. During two annual campaigns, GHL-treatment efficiently stimulated the growth of QS-degrading bacterial population of Rhodococcus erythropolis in the rhizosphere of potato plants. Analytical chemistry showed that GHL rapidly disappeared because it could be assimilated as a carbon source by R. erythropolis. Moreover, pyrosequencing of the rrs-amplicons revealed a strong modification of the structure and diversity of bacterial populations, when GHL-treated and untreated conditions were compared. This work highlighted a potential innovative strategy for stimulating the growth and root colonization of QS-degrading bacteria, which would act as biocontrol agents against plant QS-pathogens.

Proline antagonizes GABA-induced quenching of quorum-sensing in Agrobacterium tumefaciens.

Plants accumulate free L-proline (Pro) in response to abiotic stresses (drought and salinity) and presence of bacterial pathogens, including the tumor-inducing bacterium Agrobacterium tumefaciens. However, the function of Pro accumulation in host-pathogen interaction is still unclear. Here, we demonstrated that Pro antagonizes plant GABA-defense in the A. tumefaciens C58-induced tumor by interfering with the import of GABA and consequently the GABA-induced degradation of the bacterial quorum-sensing signal, 3-oxo-octanoylhomoserine lactone. We identified a bacterial receptor Atu2422, which is implicated in the uptake of GABA and Pro, suggesting that Pro acts as a natural antagonist of GABA-signaling. The Atu2422 amino acid sequence contains a Venus flytrap domain that is required for trapping GABA in human GABA(B) receptors. A constructed atu2422 mutant was more virulent than the wild type bacterium; moreover, transgenic plants with a low level of Pro exhibited less severe tumor symptoms than did their wild-type parents, revealing a crucial role for Venus flytrap GABA-receptor and relative abundance of GABA and Pro in host-pathogen interaction.

Anaesthetic management of tracheal surgery using the neodymium-yttrium-aluminium-garnet laser.

A new technique for the management of tracheal or bronchial lesions (tumours or stenosis) using the NdYAG laser and bronchoscopy, was used in 55 patients without complications. The anesthetic problems are discussed. In some circumstances the procedure may be performed using sedation and topical anaesthesia. In most, general anaesthesia was used, controlled ventilation being achieved through the bronchoscope using a modification of the Sander's injector with mixture of 50% nitrogen and oxygen to avoid the risk of fire.

[Trans-sphenoidal pituitary resections. Various problems in anesthesia and resuscitation. Study in 1,193 cases (author's transl)]. / La chirurgie trans-sphénoïdale. Problèmes posés à l'anesthésiste-réanimateur. A propos de 1193 cas.

In the present study based on 1,193 cases of trans-sphenoidal pituitary resections, the various problems involved in anaesthesia and pre and post-operative care are discussed. The anaesthetic complications are few and should be avoided through adequate preparation and supervision. The procedure is a short one (75 +/- 16 minutes) and blood loss minimal. The patients are usually young (42 +/- 4 years old), in good condition (90% were in ASA group I or II). The pre and post-operative management must follow some well defined principles, based on hormonal substitution and diagnosis and early treatment of a possible diabetes insipidus.

[Air embolism in neurosurgery. General data (author's transl)]. / Embolies gazeuses en neurochirurgie. Données générales.

Air embolism in neurosurgery, in the sitting position, are not rare. They depend on the type of surgery (more frequent in operations on the posterior fossa), on the mode of ventilation, and most of all on the criteria for detection (clinical, electrocardiographic, ultrasonic, capnographic of haemodynamia). They depend as well upon the degree of tilt, of intrathoracic and intracardiac pressures (right auricle) and of the gas mixture administered (nitrous oxide increases their size owing to its poor blood solubility). The way of entry is venous, but it is certain that they can pass into the systemic circulation (their passage through the lungs is debated). According to their size, the speed of injection and the subject's previous condition they may affect: --the lungs: precapillary pulmonary artery hypertension, and even pulmonary oedema, --the heart, mainly through the fall of venous return, --the brain anoxia, followed by cerebral oedema, leading to early neurologic disturbances and possibly severe sequellae. Those symptoms are due to the gas embolism itself, but also to microthrombi of fibrin and disturbances of vascular membrane permeability.

[Anaesthesia in microsurgery of the larynx using a carbon dioxide laser, based on 665 cases (author's transl)]. / L'anesthésie pour la microchirurgie laryngée sous laser. A propos de 665 cas.

The authors describe their experience and emphasize the risk of ignition should inflammable gases be used less the cuff should burst, and its prevention by resorting to intravenous anaesthetic agents, and ventilating the patient with air or enriched air. They describe as well their experience with the Carden tube with jet ventilation with air.

[Pharmacology of nitroglycerin (author's transl)]. / Pharmacologie de la nitroglycérine.

Nitroglycerin is a vasodilating agent by virtue of its actions on vascular smooth muscle fibers. It may be administered intravenously (using either 5 p. cent dextrose, or propylene-glycol solvant), sublingually, orally or by topical administration. It is rapidly metabolized, principally by liver. Its is not toxic. The vasodilatation that is produced is both arterial and venous and is dose-related in dog (1 microgram to 100 micrograms/kg/min). However, resistance and tachphylaxis may occur. Its principal use is for angor treatment, but it has been used for the treatment of arteriopathy of the lower limbs, biliar hypertony and arterial hypertension. It has been recently administered for the treatment of acute phase of myocardial infarction and during pre, per- and post-operative periods in cardiac surgery, neurosurgery and hip surgery, as myocardial protector or anti-hypertensive agent or hypotensive agent. The absence of toxicity and the rapid reversibility of its cardio-vascular effects which are similar to the effects of sodium nitroprusside are important reasons for its use in anesthesia and cardiac intensive care.