Neonatal kaempferol exposure attenuates impact of cerebral palsy model on neuromotor development, cell proliferation, microglia activation, and antioxidant enzyme expression in the hippocampus of rats.

OBJECTIVES: This study aims to assess the effect of neonatal treatment with kaempferol on neuromotor development, proliferation of neural precursor cells, the microglia profile, and antioxidant enzyme gene expression in the hippocampus. METHODS: A rat model of cerebral palsy was established using perinatal anoxia and sensorimotor restriction of hindlimbs during infancy. Kaempferol (1 mg/ kg) was intraperitoneally administered during the neonatal period. RESULTS: Neonatal treatment with kaempferol reduces the impact of the cerebral palsy model on reflex ontogeny and on the maturation of physical features. Impairment of locomotor activity development and motor coordination was found to be attenuated by kaempferol treatment during the neonatal period in rats exposed to cerebral palsy. Neonatal treatment of kaempferol in cerebral palsy rats prevents a substantial reduction in the number of neural precursor cells in the dentate gyrus of the hippocampus, an activated microglia profile, and increased proliferation of microglia in the sub-granular zone and in the granular cell layer. Neonatal treatment with kaempferol increases gene expression of superoxide dismutase and catalase in the hippocampus of rats submitted to the cerebral palsy model. DISCUSSION: Kaempferol attenuates the impact of cerebral palsy on neuromotor behavior development, preventing altered hippocampal microglia activation and mitigating impaired cell proliferation in a neurogenic niche in these rats. Neonatal treatment with kaempferol also increases antioxidant defense gene expression in the hippocampus of rats submitted to the cerebral palsy model.

Antibiotic-induced microbiota depletion in normally-reared adult rats mimics the neuroendocrine effects of early life stress.

Early life stress induced by maternal separation (MS) causes neuroendocrine, behavioral, and metabolic alterations that are related to gut dysbiosis. MS also increases microglial activation and decreases neurogenesis. Whether these long-term alterations are maintained or worsened in the absence of gut microbiota remains unknown. Hence, this study evaluated the effect of MS symptomatology after antibiotic-induced microbiota depletion (AIMD) in adult rats. Control and maternally separated (3 h per day from postnatal day one to 14, MS180) rats were subjected to AIMD for one month, then assessed for behavioral, metabolic, and neuroendocrine responses. Effects of MS180 and AIMD on gut microbiota were confirmed by qPCR. The data indicate that MS180 caused a passive coping strategy in the forced swimming test and decreased hippocampal neurogenesis. In addition, fasting glucose, cholesterol, and corticosterone levels increased, which correlated with a decrease in Lactobacillus spp counts in the caecum. AIMD also increased immobility in the forced swimming test, decreased hippocampal neurogenesis, and augmented corticosterone levels. However, it had no effects on glucose homeostasis or plasma lipid levels. Furthermore, the MS180-induced long-term effects on behavior and neurogenesis were not affected by microbiota depletion. Meanwhile, the metabolic imbalance was partially reversed in MS180 + AIMD rats. These results show that AIMD mimics the behavioral consequences of MS180 but may prevent metabolic imbalance, suggesting that gut dysbiosis could be part of the mechanisms involved in the maintenance of the long-term consequences of early life stress.

Bacterial and parasite co-infection in Mexican golden trout (Oncorhynchus chrysogaster) by Aeromonas bestiarum, Aeromonas sobria, Plesiomonas shigelloides and Ichthyobodo necator.

BACKGROUND: Bacterial infections are responsible of high economic losses in aquaculture. Mexican golden trout (Oncorhynchus chrysogaster) is a threatened native trout species that has been introduced in aquaculture both for species conservation and breeding for production and for which no studies of bacterial infections have been reported. CASE PRESENTATION: Fish from juvenile stages of Mexican golden trout showed an infectious outbreak in a farm in co-culture with rainbow trout (Oncorhynchus mykiss), showing external puntiform red lesions around the mouth and caudal pedunculus resembling furuncles by Aeromonas spp. and causing an accumulated mortality of 91%. Isolation and molecular identification of bacteria from lesions and internal organs showed the presence of Aeromonas bestiarum, Aeromonas sobria, Plesiomonas shigelloides and Ichthyobodo necator isolated from a single individual. All bacterial isolates were resistant to amoxicillin-clavulanic acid and cefazoline. P. shigelloides was resistant to third generation ß-lactamics. CONCLUSIONS: This is the first report of coinfection by Aeromonas bestiarum, Aeromonas sobria, Plesiomonas shigelloides and Ichthyobodo necator in an individual of Mexican golden trout in co-culture with rainbow trout. Resistance to ß-lactams suggests the acquisition of genetic determinants from water contamination by human- or livestock-associated activities.

Non-Antibiotics Strategies to Control Salmonella Infection in Poultry.

Salmonella spp. is a facultative intracellular pathogen causing localized or systemic infections, involving economic and public health significance, and remains the leading pathogen of food safety concern worldwide, with poultry being the primary transmission vector. Antibiotics have been the main strategy for Salmonella control for many years, which has allowed producers to improve the growth and health of food-producing animals. However, the utilization of antibiotics has been reconsidered since bacterial pathogens have established and shared a variety of antibiotic resistance mechanisms that can quickly increase within microbial communities. The use of alternatives to antibiotics has been recommended and successfully applied in many countries, leading to the core aim of this review, focused on (1) describing the importance of Salmonella infection in poultry and the effects associated with the use of antibiotics for disease control; (2) discussing the use of feeding-based (prebiotics, probiotics, bacterial subproducts, phytobiotics) and non-feeding-based (bacteriophages, in ovo injection, vaccines) strategies in poultry production for Salmonella control; and (3) exploring the use of complementary strategies, highlighting those based on -omics tools, to assess the effects of using the available antibiotic-free alternatives and their role in lowering dependency on the existing antimicrobial substances to manage bacterial infections in poultry effectively.

The RNA polymerase II subunit NRPB2 is required for indeterminate root development, cell viability, stem cell niche maintenance, and de novo root tip regeneration in Arabidopsis.

The RNA polymerase II drives the biogenesis of coding and non-coding RNAs for gene expression. Here, we describe new roles for its second-largest subunit, NRPB2, on root organogenesis and regeneration. Down-regulation of NRPB2 activates a determinate developmental program, which correlated with a reduction in mitotic activity, cell elongation, and size of the root apical meristem. Noteworthy, nrpb2-3 mutants manifest cell death in pro-vascular cells within primary root tips of plants grown in darkness or exposed to light, which triggers the expression of the regeneration gene marker ERF115 in neighbor cells close to damage. Auxin and stem cell niche (SCN) gene expression as well as structural analysis revealed that NRPB2 maintains SCN activity through distribution of PIN transporters in root tissues. Wild-type seedlings regenerated the root tip after excision of the QC and SCN, but nrpb2-3 mutants did not rebuild the missing tissues, and this process could be genotypified using pERF115:GFP, DR5:GFP, and pWOX5:GFP reporter constructs. The levels of reactive oxygen species increased in the mutants four days after germination and strongly decreased at later times, whereas nitric oxide accumulated as the root tip differentiates. These results show the importance of the transcriptional machinery for root organogenesis, cell viability, and regenerative capacity for reconstruction of tissues and organs upon injury.

Early life stress increases vulnerability to the sequelae of pediatric mild traumatic brain injury.

Early life stress (ELS) is a risk factor for many psychopathologies that happen later in life. Although stress can occur in cases of child abuse, studies on non-accidental brain injuries in pediatric populations do not consider the possible increase in vulnerability caused by ELS. Hence, we sought to determine whether ELS increases the effects of pediatric mild traumatic brain injury (mTBI) on cognition, hippocampal inflammation, and plasticity. Male rats were subjected to maternal separation for 180 min per day (MS180) or used as controls (CONT) during the first 21 post-natal (P) days. At P21 the rats were anesthetized with isoflurane and subjected to a mild controlled cortical impact or sham injury. At P32 the rats were injected with the cell proliferation marker bromodeoxyuridine (BrdU, 500 mg/kg), then evaluated for spatial learning and memory in a water maze (P35-40) and sacrificed for quantification of Ki67+, BrdU+ and Iba1+ (P42). Neither MS180 nor mTBI impacted cognitive outcome when provided alone but their combination (MS180 + mTBI) decreased spatial learning and memory relative to Sham controls (p < .01). mTBI increased microglial activation and affected BrdU+ cell survival in the ipsilateral hippocampus without affecting proliferation rates. However, only MS180 + mTBI increased microglial activation in the area adjacent to the injury and the contralateral CA1 hippocampal subfield, and decreased cell proliferation in the ipsilateral neurogenic niche. Overall, the data show that ELS increases the vulnerability to the sequelae of pediatric mTBI and may be mediated by increased neuroinflammation.

Physiological and Anatomical Mechanisms in Wheat to Cope with Salt Stress Induced by Seawater.

Two pot experiments were conducted in a greenhouse to examine 14C fixation and its distribution in biochemical leaf components, as well as the physiological and anatomical adaptability responses of wheat (Triticum aestivum L.) grown with seawater diluted to 0.2, 3.0, 6.0, and 12.0 dS m-1. The results showed significant reductions in chlorophyll content, 14C fixation (photosynthesis), plant height, main stem diameter, total leaf area per plant, and total dry weight at 3.0, 6.0, and 12.0 dS m-1 seawater salt stress. The 14C loss was very high at 12.0 ds m-1 after 120 h. 14C in lipids (ether extract) showed significant changes at 12.0 dS m-1 at 96 and 120 h. The findings indicated the leaf and stem anatomical feature change of wheat plants resulting from adaptation to salinity stress. A reduction in the anatomical traits of stem and leaf diameter, wall thickness, diameter of the hollow pith cavity, total number of vascular bundles, number of large and small vascular bundles, bundle length and width, thickness of phloem tissue, and diameter of the metaxylem vessel of wheat plants was found. In conclusion, salt stress induces both anatomical and physiological changes in the stem and leaf cells of wheat, as well as the tissues and organs, and these changes in turn make it possible for the plants to adapt successfully to a saline environment.

Agricultural reuse of municipal wastewater through an integral water reclamation management.

The DESERT-prototype, a state-of-the-art compact combination of water treatment technologies based on filtration and solar-based renewable energy, was employed to reclaim water for agricultural irrigation. Water reclaimed through the DESERT-prototype (PW) from a secondary effluent of a wastewater treatment plant, as well as conventional irrigation water (CW) and the secondary effluent (SW) itself, were employed to cultivate baby romaine lettuces in a greenhouse in Murcia (Spain), by means of drip and sprinkler irrigation methods, thus establishing six treatments. Assessments of physicochemical and microbiological quality of irrigation water, as well as agronomic and microbiological quality of crops from all treatments, showed that results associated to PW complied in all cases with relevant standards and guidelines. In contrast, results linked to SW and CW presented certain non-compliance cases of water and crop microbiological quality. These assessments lead to conclude that the DESERT-prototype is an appropriate technology for safe water reclamation oriented to agricultural production, that can be complemented by a proper irrigation method in reaching safety targets.

P-wave and interatrial block: New predictor for atrial high rate episodes in patients with cardiac implantable electronic devices.

BACKGROUND: The presence of interatrial block (IAB) is associated with the development of atrial fibrillation (AF). The aim of this study was to determine whether P-wave duration and presence of IAB before the implantation of a cardiac implantable electronic device (CIED) are associated with the presence of atrial high rate episodes (AHRE), during long-term follow-up. METHODS: 380 patients (57% men; 75 ± 10 years) were included. IAB was defined according to the International Consensus Criteria. AHRE was defined as an episode of atrial rate ≥225 beats/min with a minimum duration of 5 minutes. RESULTS: Documented paroxysmal AF before the implantation was present in 24% of the patients; 80% had hypertension and 32% structural heart disease. Mean P-wave duration was 123 ± 23 ms, and 39% of the patients had IAB (32% partial, 7% advanced). After a mean follow-up of 18 ± 12 months, 33% of the patients presented AHRE. Patients with AHRE had a P-wave duration significantly longer (130 ± 24 ms vs 119 ± 21 ms; P < 0.001) and a greater prevalence of IAB (53% vs 32%; P < 0.001). In a multivariate analysis, predictors of AHRE were: IAB (odds ratio [OR] 2.1; 95% confidence interval [CI] [1.3-3.4], P < 0.001) and previous paroxysmal AF (OR 2.6; 95% CI [1.5-4.3], P < 0.001). In patients without previous AF, the presence of IAB was also a significant predictor of AHRE (OR 3.1; 95% CI [1.8-5.5], P < 0.001). CONCLUSIONS: IAB is a strong predictor of AHRE in patients with CIED. This finding is independent of the presence of prior paroxysmal AF.

Prevalence of Human Papillomavirus in Women from the State of Michoacan, Mexico, Showed High Frequency of Unusual Virus Genotypes.

BACKGROUND: Human papillomaviruses (HPVs), the leading cause of cervical cancer, are distributed worldwide, with high prevalence in developing countries. OBJECTIVE: The objective of the study is to know the prevalence and genotypes of HPV in women from the state of Michoacán and the Women's Hospital in Morelia, Michoacán. MATERIALS AND METHODS: Cervical smear samples (159,288) were subjected to HPV detection by hybrid capture 2. A subsample of 484 patients from the Women's Hospital was studied by Papanicolaou test and linear array HPV genotyping, and when positive, patients were also examined by colposcopy and histopathology. RESULTS: The overall prevalence for HPV in Michoacán State was 7.74%; 7.11% in 2009, 6.46% in 2010, 9.58% in 2011, and 8.43% in 2012. The highest prevalence was found in the age groups < 25 and 25-34 years. The prevalence at the Women's Hospital was 8.51%. Cytological examination revealed normal cytology in 64.44% of samples, 26.66 % with low-grade and 8.88 % with high-grade squamous intraepithelial lesion (HSIL). However, by colposcopy, normal tissue appearance was found only in 26.66%; 51% were reclassified as low-grade squamous intraepithelial lesion, 17.77% as HSIL, and in 4.4% atrophy was observed. The most prevalent genotype in single infections was HPV59, followed by HPV51 and HPV45. Double infections occurred with the following genotypes: 52-53, 51-59, 61-67, 66-11, 16-62, 53-62, 59-CP6108, 45-66, and 45-51. Triple infections were identified as: 6-31-39, 51-59-62, 51-62-81, 54-55-59, 16-58-71, and 16-59-62. CONCLUSIONS: The prevalent genotype found among women from Michoacán, HPV59, was different to the rest of the country. The high prevalence of HPV59 could be due to cases imported to Michoacán by agricultural workers migrating to the USA or may be associated to ethnicity differences. Implications of this finding for immunization programs should be explored.

The CSL proteins, versatile transcription factors and context dependent corepressors of the notch signaling pathway.

The Notch signaling pathway is a reiteratively used cell to cell communication pathway that triggers pleiotropic effects. The correct regulation of the pathway permits the efficient regulation of genes involved in cell fate decision throughout development. This activity relies notably on the CSL proteins, (an acronym for CBF-1/RBPJ-κ in Homo sapiens/Mus musculus respectively, Suppressor of Hairless in Drosophila melanogaster, Lag-1 in Caenorhabditis elegans) which is the unique transcription factor and DNA binding protein involved in this pathway. The CSL proteins have the capacity to recruit activation or repression complexes according to the cellular context. The aim of this review is to describe the different co-repressor proteins that interact directly with CSL proteins to form repression complexes thereby regulating the Notch signaling pathway in animal cells to give insights into the paralogous evolution of these co-repressors in higher eumetazoans and their subsequent effects at developmental processes.

Safety assessment of greenhouse hydroponic tomatoes irrigated with reclaimed and surface water.

The impact of reclaimed and surface water on the microbiological safety of hydroponic tomatoes was assessed. Greenhouse tomatoes were irrigated with reclaimed and surface water and grown on two hydroponic substrates (coconut fiber and rock wool). Water samples (n=208) were taken from irrigation water, with and without the addition of fertilizers and drainage water, and hydroponic tomatoes (n=72). Samples were analyzed for indicator microorganisms, generic Escherichia coli and Listeria spp., and pathogenic bacteria such as Salmonella spp. and Shiga-toxigenic E. coli (STEC), using multiplex real-time PCR (RT-PCR) after enrichment. The correlation between climatological parameters such as temperature and the levels of microorganisms in water samples was also determined. In irrigation water, generic E. coli counts were higher in reclaimed than in surface water whereas Listeria spp. numbers increased after adding the fertilizers in both water sources. In drainage water, no clear differences in E. coli and Listeria numbers were observed between reclaimed and surface water. No positive samples for STEC were found in irrigation water. Presumptive positives for Salmonella spp. were found in 7.7% of the water samples and 62.5% of these samples were reclaimed water. Salmonella-positive samples by RT-PCR could not be confirmed by conventional methods. Higher concentrations of E. coli were associated with Salmonella-presumptive positive samples. Climatological parameters, such as temperature, were not correlated with the E. coli and Listeria spp. counts. Tomato samples were negative for bacterial pathogens, while generic E. coli and Listeria spp. counts were below the detection limit. The prevalence of presumptive Salmonella spp. found in irrigation water (reclaimed and surface water) was high, which might present a risk of contamination. The absence of pathogens on greenhouse hydroponic tomatoes indicates that good agricultural practices (GAP) were in place, avoiding the microbial contamination of the fruit.

Evaluation of DNA extraction methods of rumen microbial populations.

The dynamism of microbial populations in the rumen has been studied with molecular methods that analyze single nucleotide polymorphisms of ribosomal RNA gene fragments (rDNA). Therefore DNA of good quality is needed for this kind of analysis. In this work we report the evaluation of four DNA extraction protocols (mechanical lysis or chemical lysis with CTAB, ethylxanthogenate or DNAzol(®)) from ruminal fluid. The suitability of two of these protocols (mechanical lysis and DNAzol(®)) was tested on single-strand conformation polymorphism (SSCP) of rDNA of rumen microbial populations. DNAzol(®) was a simple method that rendered good integrity, yield and purity. With this method, subtle changes in protozoan populations were detected in young bulls fed with slightly different formulations of a supplement of multinutritional blocks of molasses and urea. Sequences related to Eudiplodinium maggi and a non-cultured Entodiniomorphid similar to Entodinium caudatum, were related to major fluctuating populations in an SSCP assay.

Structural analysis of point mutations in the Hairless gene and their association with the activity of the Hairless protein.

The Notch signaling pathway (NSP) is an important intercellular communication mechanism that regulates embryo development and adult physiological functions. The Hairless (H) protein is a powerful antagonist of the NSP by its interaction with the Suppressor of Hairless (Su[H]) protein, recruiting the corepressors Gro and CtBP. In the present work, we examined the role of several important amino acids in different H protein domains analyzing four mutant lines of Drosophila melanogaster. The mutant alleles were evaluated by single-strand conformational polymorphism (SSCP) analysis and we located mutated regions at different positions along the sequence of the Hairless gene.

Changes in photosynthetic parameters and antioxidant activities following heat-shock treatment in tomato plants.

Seedlings of two tomato genotypes, Lycopersicon esculentum Mill. var. Amalia and the wild thermotolerant type Nagcarlang, were grown under a photoperiod of 16 h light at 25°C and 8 h dark at 20°C. At the fourth true leaf stage, a group of plants were exposed to a heat-shock temperature of 45°C for 3 h, and measurements of chlorophyll fluorescence, gas-exchange characteristics, dark respiration and oxidative and antioxidative parameters were made after releasing the stress. The heat shock induced severe alterations in the photosynthesis of Amalia that seem to mitigate the damaging impact of high temperatures by lowering the leaf temperature and maintaining stomatal conductance and more efficient maintenance of antioxidant capacity, including ascorbate and glutathione levels. These effects were not evident in Nagcarlang. In Amalia plants, a larger increase in dark respiration also occurred in response to heat shock and the rates of the oxidative processes were higher than in Nagcarlang. This suggests that heat injury in Amalia may involve chlorophyll photooxidation mediated by activated oxygen species (AOS) and more severe alterations in the photosynthetic apparatus. All these changes could be related to the more dramatic effect of heat shock seen in Amalia than in Nagcarlang plants.

Sap flow, gas exchange, and hydraulic conductance of young apricot trees growing under a shading net and different water supplies.

The experiment was carried out in a research field near Murcia, Spain, over a 3-week period between September 26 and October 16, 2000. Sixteen trees were used in the experiment, eight of which were placed under a rectangular shading net, while the other eight were maintained in the open air. Trees were irrigated once per day and, after October 5th, water was witheld from eight trees (four shaded and four unprotected for 5 days). The leaf stomatal conductance and the photosynthesis rates were higher in the shaded trees than in the exposed plants, probably because the leaf water potential was lower in the unshaded plants. This higher leaf conductance partially compensated for the effect of low radiation on transpiration, and the reduction of daily sap flow registered in shaded trees was only around 10-20%. The net also affected trunk diameter changes, with the shaded trees showing lower values of maximum daily shrinkage. Soil water deficit and high radiation had a similar effect on plant water parameters, lowering leaf water potential, leaf stomatal conductance, and the photosynthesis rate. The effects of both conditions were accumulative and so the exposed water-stressed plants showed the lowest values of total hydraulic resistance and water use efficiency, while the shaded well-irrigated trees registered the highest values for both parameters. For this reason, we think that net shading could be extended to apricot culture in many areas in which irrigation water is scarce and insolation is high.

High temperature effects on photosynthetic activity of two tomato cultivars with different heat susceptibility.

The functional activities of the photosynthetic apparatus of two tomato cultivars of different thermotolerance were investigated after a short period of high temperature treatment. Seedlings of two tomato genotypes, Lycopersicon esculentum var. Campbell-28 and the wild thermotolerant Nagcarlang, were grown under a photoperiod of 16h at 25 degrees C and dark period of 8h at 20 degrees C. At the fourth true leaf stage, a group of plants was exposed to heat stress of 45 degrees C for 2 h. The heat shock treatment caused important reductions of the net photosynthetic rate (Pn) of Campbell-28 plants due to non-stomatal components. These non-stomatal effects were not evident in Nagcarlang-treated plants. This reduction in the CO2 assimilation rate observed in Campbell-28 was generated by affections in the Calvin cycle and also in the PSII functioning. No changes in these parameters were observed in the thermotolerant genotype after the stress. Injury to the plasma membrane because of the heat stress was evident only in the Campbell-28 genotype. Heat led to a sun-type adaptation response of the photosynthesis pigment apparatus for the Nagcarlang genotype, but not for Campbell-28, and thus an increase in chlorophyll a/b ratio and a decrease in chlorophyll/carotenoid ratio were shown in Nagcarlang stressed plants.

Effects of irrigation and air humidity preconditioning on water relations, growth and survival of Rosmarinus officinalis plants during and after transplanting.

The effect of different irrigation and air humidity conditioning treatments on the morphological and physiological responses of Rosmarinus officinalis in nursery conditions was investigated in order to evaluate the degree of hardening resulting from these conditions. Rosmarinus officinalis seedlings were pot-grown during 4 months in two greenhouses (nursery period), in which two irrigation treatments were used (control and deficit). In one of these greenhouses, air humidity was controlled using a dehumidifying system (low humidity), in the other greenhouse the air conditions were not artificially modified (control humidity). After the nursery period, the plants of all treatments were transplanted and well watered (100% water holding capacity for 1 month, transplanting period). After this period, they received no water (establishment period). At the end of the nursery period it was seen that deficit irrigation had altered the morphology of the R. officinalis plants by reducing plant height, stem diameter, leaf area, total dry weight, and root length, while humidity influenced the parameters related with plant water relations. Low air humidity and deficit irrigation-induced tissue dehydration and lower stomatal conductance values (gs). The plants subjected to deficit irrigation developed leaf osmotic adjustment, which was maintained during the transplanting period. At that time, the plants that had been exposed to deficit irrigation and low humidity showed efficient stomatal regulation (lower gs values). After transplanting and during the establishment period, these plants showed a better water status (higher psil and gs values). Their post-planting survival rate improved as a result of acclimation processes.

Variations in water status, gas exchange, and growth in Rosmarinus officinalis plants infected with Glomus deserticola under drought conditions.

The influence of the arbuscular mycorrhizal fungus Glomus deserticola on the water relations, gas exchange parameters, and vegetative growth of Rosmarinus officinalis plants under water stress was studied. Plants were grown with and without the mycorrhizal fungus under glasshouse conditions and subjected to water stress by withholding irrigation water for 14 days. Along the experimental period, a significant effect of the fungus on the plant growth was observed, and under water stress, mycorrhizal plants showed an increase in aerial and root biomass compared to non-mycorrhizal plants. The decrease in the soil water potential generated a decrease in leaf water potential (psi(l)) and stem water potential (psi(x)) of mycorrhizal and non-mycorrhizal plants, with this decrease being lower in mycorrhizal water-stressed plants. Mycorrhization also had positive effects on the root hydraulic conductivity (Lp) of water stressed plants. Furthermore, mycorrhizal-stressed plants showed a more important decrease in osmotic potential at full turgor (psi(os)) than did non-mycorrhizal-stressed plants, indicating the capacity of osmotic adjustment. Mycorrhizal infection also improved photosynthetic activity (Pn) and stomatal conductance (g(s)) in plants under water stress compared to the non-mycorrhizal-stressed plants. A similar behaviour was observed in the photochemical efficiency of PSII (Fv/Fm) with this parameter being lower in non-mycorrhizal plants than in mycorrhizal plants under water stress conditions. In the same way, under water restriction, mycorrhizal plants showed higher values of chlorophyll content than did non-mycorrhizal plants. Thus, the results obtained indicated that the mycorrhizal symbiosis had a beneficial effect on the water status and growth of Rosmarinus officinalis plants under water-stress conditions.

Molecular characterization of Bacillus thuringiensis strains from Argentina.

Bacillus thuringiensis INTA 7-3, INTA 51-3, INTA Mo9-5 and INTA Mo14-4 strains were obtained from Argentina and characterized by determination of serotype, toxicity, plasmid composition, insecticidal gene content ( cry and vip ) and the cloning of the single- vip3A gene of the INTA Mo9-5 strain. The serotype analysis identified the serovars tohokuensis and darmstadiensis for the INTA 51-3 and INTA Mo14-4 strains, respectively, whereas the INTA Mo9-5 strain was classified as "autoagglutinated". In contrast to the plasmid patterns of INTA 7-3, INTA 51-3 and INTA Mo9-5 (which were similar to B. thuringiensis HD-1 strain), strain INTA Mo14-4 showed a unique plasmid array. PCR analysis of the four strains revealed the presence of cry genes and vip3A genes. Interestingly, it was found that B. thuringiensis 4Q7 strain, which is a plasmid cured strain, contained vip3A genes indicating the presence of these insecticidal genes in the chromosome. Bioassays towards various lepidopteran species revealed that B. thuringiensis INTA Mo9-5 and INTA 7-3 strains were highly active. In particular, the mean LC(50) obtained against A. gemmatalis larvae with the INTA Mo9-5 and INTA 7-3 strains were 7 (5.7-8.6) and 6.7 (5.6-8.0) ppm, respectively. The INTA Mo14-4 strain was non-toxic and strain INTA 51-3 showed only a weak larvicidal activity.