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The functional role of the insect pathogen fungus Metarhizium anisopliae strain NCAIM 362 against the white grub cockchafer (Melolontha melolontha) larvae and southern European marshland pyralid (Duponchelia fovealis) larvae in a sweet potato cultivation was tested under greenhouse conditions. As a positive control, the effect of the same strain of M. anisopliae were compared with the effect of the synthetic insecticide alpha-cypermethrin. According to the results, alpha-cypermethrin was more effective against both the Coleoptera and Lepidoptera larvae, a lower number of surviving individuals as well as less damaged tubers were detected after the chemical treatment, compared with M. anisopliae.

Ticks can be a major concern for humans and animals alike through the transmission of various viral and bacterial diseases. Ticks have also developed tolerance to several active compounds due to intensive insecticide treatments. The excess use of insecticides against ticks worldwide is mostly environmental contamination rather than effective control. Thus, information about the useful chemicals is essential. Dermacentor marginatus and Dermacentor reticulatus individuals were tested, being collected in Transylvania, near the city of Târgu-Mureş, Romania. The experiment was performed under laboratory conditions in two replicates, using a total of 420 ticks and 19 insecticides. The best results were achieved with alpha-cypermethrin and pyrethrin.

Oana Codruta Miloicov Bacean,1 Ovidiu Fira-Mladinescu,2 Salomeia Putnoky,1 Ioana Tuta Sas,1 Petrescu Cristina,1 Radu Bagiu,1 Iulia Cristina Bagiu,3 Oana Suciu,1 Corneluta Fira-Mladinescu,1 Brigitha Vlaicu1 1Discipline of Hygiene, Department 14 Microbiology, “Victor Babes” University of Medicine and Pharmacy Timisoara, Center for Studies in Preventive Medicine, Timisoara, Timis, Romania; 2Pneumology Discipline, Department 13th Infectious Diseases, “Victor Babes” University of Medicine and Pharmacy Timisoara, Center for Studies in Preventive Medicine, Timisoara, Timis, Romania; 3Discipline of Microbiology, Department 14 Microbiology, “Victor Babes” University of Medicine and Pharmacy Timisoara, Center for Studies in Preventive Medicine, Timisoara, Timis, RomaniaCorrespondence: Oana Codruta Miloicov Bacean Dumbravita, str Fiziii, nr.8, Timis County, RomaniaTel +40745170879Email baceano@gmail.comPurpose: The aims of this study were to evaluate high school pupils’ relationships with their parents, schoolmates, and friends, and the presence of health-related risk behaviors among the family members and friends on the pupils’ smoking and binge drinking status.Materials and Methods: The study was a cross-sectional population-based study that involved a questionnaire, and the sample involved 2908 pupils at high schools in Timis county, Romania. In our study, the following factors related to family members and friends and the living environment were identified. First, there were issues with the relationships with parents among some pupils, with 23.2% being neither satisfied nor unsatisfied and 4.9% being unsatisfied. Second, there were also issues regarding some pupils’ satisfaction with the financial situation of their family, with 21.8% being neither satisfied nor unsatisfied and 16.4% being unsatisfied. An odds ratio (OR) is a statistic that quantifies the strength of the association between two events.Results: There were 1495 (51.5%) girls and 1407 (48.5%) boys aged 14– 20 years, and 79.2% came from organized families. Among the pupils, 71.1% reported that they were satisfied with their relationship with their parents, while 4.9% were unsatisfied. Additionally, 46.2% were satisfied with the financial status of their families, while 16.4% were unsatisfied. Moreover, 59.2% were satisfied with their relationship with their schoolmates and 80.2% were satisfied with their relationship with their friends, while only 1.8% were unsatisfied with their relationship with their friends. The logistic regression model of the pupils’ smoking status identified two moderate predictive factors: this was defined “> 1” friends who smoke (odds ratio [OR]=2.875) and smoking status of siblings (OR=2.222). The major predictive factor in the logistic regression model of occasional alcohol consumption in high quantities by pupils was male gender (OR=5.148).Conclusion: The predictive model of pupils’ smoking status identified the following moderate predictors: high number of friends who smoke and smoking status of siblings. The major predictor in the model of occasional alcohol consumption in high quantities by pupils was male gender.Keywords: teenagers, pupils, family, entourage, health risk

Negative emotions and chronic stress trigger abnormal compensatory behaviors known as emotional eating (EE). EE is a well-known mediator for increased body mass index and weight gain. Our aim was to analyze the factor structure and validity and reliability of the Emotional Eater Questionnaire (EEQ) in a sample of 200 Romanian adults with excess weight. Principal component analysis (PCA) was used to assess the construct validity. The mindful eating questionnaire (MEQ) was used to test concurrent validity. Cronbach’s alpha and Spearman correlations were used to analyze internal and external reliability. The socio-demographic characteristics were used as factors for convergent validity. PCA revealed the existence of three major factors, disinhibition, type of food, and guilt, which accounted for 64.9% of the variance. Concurrent validity showed medium to large associations with MEQ (r = 0.650; p < 0.001) and a large association with the emotional subscale of MEQ (r = 0.732; p < 0.001). Reliability was adequate with Cronbach’s alfa = 0.841 and ICC = 0.775. In a multivariate model, the highest contribution to the EE score was the age (beta = −0.327), followed by feminine gender (beta = 0.321), high levels of perceived stress (beta = 0.215), BMI (beta = 0.184) and lower perceived health status (beta = 0.184). The Romanian version of the EEQ is a reliable and valid tool for measuring emotional eating in adults with excess weight.

In Romania, one in four children has excess weight. Because childhood obesity is a sensitive topic, many healthcare professionals find it difficult to discuss children's excess weight with parents. This study aims to identify barriers and facilitators in childhood obesity-related communication, as perceived by healthcare professionals in Romania. As part of the STOP project, healthcare professionals (family physicians, pediatricians, and dieticians) who treat children with excess weight were invited to a telephone interview. The semi-structured questions were translated from a questionnaire previously used at the Swedish study site of the STOP project. Interviews were transcribed and then used for thematic analysis. Fifteen doctors and three dieticians (16 females and 2 males), with average 18.2 ± 10.1 years of experience, were interviewed. Four main themes were identified. Professionals reported that when children began experiencing obesity-related stigma or comorbidities, this became the tipping point of weight excess, where parents felt motivated to begin treatment. Barriers in communication were part of several layers of distrust, recognized as tension between professionals and caregivers due to conflicting beliefs about excess weight, as well as lack of trust in medical studies. Most respondents felt confident using models of good practice, consisting of a gentle approach and patient-centered care. Nonetheless, professionals noted systemic barriers due to a referral system and allocation of clinical time that hinder obesity treatment. They suggested that lack of specialized centers and inadequate education of healthcare professional conveys the system does not prioritize obesity treatment and prevention. The interviewed Romanian doctors and dieticians identified patient-centered care as key to treating children with obesity and building trust with their caregivers. Howevertheir efforts are hindered by healthcare system barriers, including the lack of specialized centers, training, and a referral system. The findings therefore suggest that, to improve childhood obesity prevention and treatment, systemic barriers should be addressed.Trial Registration:ClinicalTrials.gov, NCT03800823; 11 Jan 2019.

In depth understanding of the dietary patterns of individuals with obesity is needed in practice and research, in order to support dietitians and physicians in the design and implementation of nutritional management. We aimed to analyze the consistency of energy, macro-, and micronutrient reported intakes in four non-consecutive 24-h dietary recalls from 388 adults with obesity using information collected in the NutriGen Study (ClinicalTrials.gov, NCT02837367). Significant decreases were identified for reported energy and several, macro- and micronutrient intakes, between the first and subsequent 24-h recalls. Significant differences of reported intakes were identified in sensitivity analyses, suggesting that the first recall (also the only one performed on site, face-to-face) might be a point of bias. A comparison of the differences in intakes between weekend and weekday, after adjustment for false discovery rate were non-statistically significant either in male, females, or in total. To overcome this potential bias, studies should be carefully conducted, starting from the design phase, through to the analysis and interpretation phases of the study. Prior to averaging specific intakes across all sessions of reporting, a preliminary analysis must be conducted to identify if a certain time point had significant differences from all other time points and overview potential sources of bias: reporting bias, training bias, or behavioral changes could be responsible for such differences.

The effect of fungal entomopathogen M. anisopliae strain NCAIM 362 against M. melolontha larvae in sweet potato was tested under open field conditions when crop management included compost supply and soil cover (agro-foil or agro-textile). Additionally, the effect of M. anisopliae same strain against M. melolontha was compared with the effect of α-cypermethrin under greenhouse conditions. Soil microbial community using Illumina sequencing and soil biological activity were tested as possible parameter influencing M. anisopliae effect. According to the results, compost supply and textile cover may enhance the effectiveness of M. anisopliae under open field conditions, while no effect of fungal treatment was detected under greenhouse conditions. Even if soil parameters (chemical composition, bacterial, and biological activity) were identical, the effect of α-cypermethrin against M. melolontha larvae was significant: lower ratio of larval survival and less damaged tubers were detected after the chemical treatment. Our results suggest that M. anisopliae strain NCAIM 362 is not effective to control M. melolontha larvae, further pieces of research are needed to test other species of the Metarhizium genus to find an effective agent for sustainable pest control in sweet potato.

Fungal entomopathogens are gaining increasing attention as alternatives to chemical control of arthropod pests, and the literature on their use under different conditions and against different species keeps expanding. Our review compiles information regarding the entomopathogenic fungal species Metarhizium flavoviride (Gams and Rozsypal 1956) (Hypocreales: Clavicipitaceae) and gives account of the natural occurrences and target arthropods that can be controlled using M. flavoviride. Taxonomic problems around M. flavoviride species sensu lato are explained. Bioassays, laboratory and field studies examining the effect of fermentation, culture regimes and formulation are compiled along with studies on the effect of the fungus on target and non-target organisms and presenting the effect of management practices on the use of the fungus. Altogether, we provide information to help conducting basic studies, and by pointing out relatively uncharted territories, help to set new research areas.

Rhizobial surface polysaccharides, including capsular polysaccharides (KPS), are involved in symbiotic infection. The rkp-3 locus of Sinorhizobium meliloti 41 is responsible for the production of pseudaminic acid, one of the components of the KR5 antigen, a strain-specific KPS. We have extended the sequence determination and genetic dissection of the rkp-3 region to clarify the structure and function of the rkpY gene and to identify additional rkp genes. Except for rkpY, no other genes were found where mutation affected the KPS structure and symbiosis. These mutants show a unique phenotype producing a low molecular weight polysaccharide (LMW PS). Creating double mutants, we have shown that biosynthesis genes of the KR5 antigen except rkpZ are not necessary for the production of this LMW PS. Polysaccharide analysis of genetically modified strains suggests that rkpY has pleiotropic effects on polysaccharide production. It directs KPS synthesis to the KR5 antigen and influences lipo-oligo 3-deoxy-d-manno-2 octulosonic acid (Kdo) production in S. meliloti 41. In addition, rkpY suppresses the lipo-oligoKdo production when it is introduced into S. meliloti 1021.

The rkp-3 region is indispensable for capsular polysaccharide (K antigen) synthesis in Sinorhizobium meliloti Rm41. Strain Rm41 produces a K antigen of strain-specific structure, designated as the KR5 antigen. The data in this report show that the rkp-3 gene region comprises 10 open reading frames involved in bacterial polysaccharide synthesis and export. The predicted amino acid sequences for the rkpL-Q gene products are homologous to enzymes involved in the production of specific sugar moieties, while the putative products of the rkpRST genes show a high degree of similarity to proteins required for transporting polysaccharides to the cell surface. Southern analysis experiments using gene-specific probes suggest that genes involved in the synthesis of the precursor sugars are unique in strain Rm41, whereas sequences coding for export proteins are widely distributed among Sinorhizobium species. Mutations in the rkpL-Q genes result in a modified K antigen pattern and impaired symbiotic capabilities. On this basis, we suggest that these genes are required for the production of the KR5 antigen that is necessary for S. meliloti Rm41 exoB (AK631)-alfalfa (Medicago sativa) symbiosis.

Plants have evolved the ability to distinguish between symbiotic and pathogenic microbial signals. However, potentially cooperative plant-microbe interactions often abort due to incompatible signaling. The Nodulation Specificity 1 ( NS1 ) locus in the legume Medicago truncatula blocks tissue invasion and root nodule induction by many strains of the nitrogen-fixing symbiont Sinorhizobium meliloti . Controlling this strain-specific nodulation blockade are two genes at the NS1 locus, designated NS1a and NS1b , which encode malectin-like leucine-rich repeat receptor kinases. Expression of NS1a and NS1b is induced upon inoculation by both compatible and incompatible Sinorhizobium strains and is dependent on host perception of bacterial nodulation (Nod) factors. Both presence/absence and sequence polymorphisms of the paired receptors contribute to the evolution and functional diversification of the NS1 locus. A bacterial gene, designated rns1 , is required for activation of NS1 -mediated nodulation restriction. rns1 encodes a type I-secreted protein and is present in approximately 50% of the nearly 250 sequenced S. meliloti strains but not found in over 60 sequenced strains from the closely related species Sinorhizobium medicae . S. meliloti strains lacking functional rns1 are able to evade NS1 -mediated nodulation blockade.

Genes encoding the tail proteins of the temperate phage 16-3 of the symbiotic nitrogen-fixing bacterium Sinorhizobium meliloti 41 have been identified. First, a new host range gene, designated hII, was localized by using missense mutations. The corresponding protein was shown to be identical to the 85-kDa tail protein by determining its N-terminal sequence. Electron microscopic analysis showed that phage 16-3 possesses an icosahedral head and a long, noncontractile tail characteristic of the Siphoviridae. By using a lysogenic S. meliloti 41 strain, mutants with insertions in the putative tail region of the genome were constructed and virion morphology was examined after induction of the lytic cycle. Insertions in ORF017, ORF018a, ORF020, ORF021, the previously described h gene, and hII resulted in uninfectious head particles lacking tail structures, suggesting that the majority of the genes in this region are essential for tail formation. By using different bacterial mutants, it was also shown that not only the RkpM and RkpY proteins but also the RkpZ protein of the host takes part in the formation of the phage receptor. Results for the host range phage mutants and the receptor mutant bacteria suggest that the HII tail protein interacts with the capsular polysaccharide of the host and that the tail protein encoded by the original h gene recognizes a proteinaceous receptor.

Rhizobial surface polysaccharides, including capsular polysaccharides (KPS), are involved in symbiotic infection. The rkp-3 locus of Sinorhizobium meliloti 41 is responsible for the production of pseudaminic acid, one of the components of the KR5 antigen, a strain-specific KPS. We have extended the sequence determination and genetic dissection of the rkp-3 region to clarify the structure and function of the rkpY gene and to identify additional rkp genes. Except for rkpY, no other genes were found where mutation affected the KPS structure and symbiosis. These mutants show a unique phenotype producing a low molecular weight polysaccharide (LMW PS). Creating double mutants, we have shown that biosynthesis genes of the KR5 antigen except rkpZ are not necessary for the production of this LMW PS. Polysaccharide analysis of genetically modified strains suggests that rkpY has pleiotropic effects on polysaccharide production. It directs KPS synthesis to the KR5 antigen and influences lipo-oligo 3-deoxy-d-manno-2 octulosonic acid (Kdo) production in S. meliloti 41. In addition, rkpY suppresses the lipo-oligoKdo production when it is introduced into S. meliloti 1021.

The pha1 gene cluster (pha1A'-G) of Sinorhizobium meliloti has previously been characterized as a necessary component for proper invasion into plant root tissue. It has been suggested to encode a multi-subunit K(+)/H(+) antiporter, since mutations in the pha1 region rendered S. meliloti cells sensitive to K(+) and alkali, and because there is high amino acid sequence similarity to previously characterized multi-subunit cation/H(+) antiporters (Mrp antiporters). However, the detailed transport properties of the Pha1 system are yet to be determined. Interestingly, most of the Mrp antiporters are highly selective for Na(+), unlike the Pha1 system. Here, we report the functional expression of the Pha1 system in Escherichia coli and the measurement of cation/H(+) antiport activity. We showed that the Pha1 system is indeed a K(+)/H(+) antiporter with a pH optimum under mildly alkaline conditions. Moreover, we found that the Pha1 system can transport Na(+); this was unexpected based on previous phenotypic analyses of pha1 mutants. Furthermore, we demonstrated that the cation selectivity of the Pha1 system was altered when the pH was lowered from the optimum. The downregulation of Na(+)/H(+) and K(+)/H(+) antiport activities upon acidic shift appeared to occur via different processes, which might indicate the presence of distinct mechanisms for the regulation of the K(+)/H(+) and Na(+)/H(+) antiport activities of the Pha1 system.

The strain-specific capsular polysaccharide KR5 antigen of Sinorhizobium meliloti 41 is required both for invasion of the symbiotic nodule and for the adsorption of bacteriophage 16-3. In order to know more about the genes involved in these events, bacterial mutants carrying an altered phage receptor were identified by using host range phage mutants. A representative mutation was localized in the rkpM gene by complementation and DNA sequence analysis. A host range phage mutant isolated on these phage-resistant bacteria was used to identify the h gene, which is likely to encode the tail fiber protein of phage 16-3. The nucleotide sequences of the h gene as well as a host range mutant allele were also established. In both the bacterial and phage mutant alleles, a missense mutation was found, indicating a direct contact between the RkpM and H proteins in the course of phage adsorption. Some mutations could not be localized in these genes, suggesting that additional components are also important for bacteriophage receptor recognition.