Prevalence of thermotolerant species of Campylobacter and their biotypes in children and domestic birds and dogs in Southern Chile

The prevalence of thermotolerant Campylobacter in diarrhoeic and healthy children as well as in dogs, hens, ducks and pigeons was determined in Southern Chile. Campylobacter were found in 34.5% of the faecal samples examined. The isolation rate of Campylobacter in diarrhoeic and healthy children was 16.3% and 6.4% respectively. Despite C.jejuni was always more frequent than C. coli, the latter was isolated with a high frequency (29%) from patients with diarrhoea. C.jejuni and C. coli biotypes I and II were found in healthy and diarrhoeic children and were predominant in all the animals species studied. This may point out towards the possible origin of strains infecting children.

Genetic diversity among proso millet (Panicum miliaceum) biotypes assessed by AFLP technique

The Amplified Fragment Length Polymorphism (AFLP) technique was used to access genetic diversity between three domestic and nine wild proso millet biotypes from the United States and Canada. Eight primer combinations detected 39 polymorphic DNA fragments, with the genetic distance estimates among biotypes ranging from 0.02 to 0.04. Colorado-Weld County black seeded and Wyoming-Platte County were the most distinct biotypes according to the dissimilarity level. A UPGMA cluster analysis revealed two distinct groups of proso millet without any geographic association. Six weed biotypes exhibiting some characters of cultivated plants were grouped together with domesticated biotypes of proso millet while the three typical wild phenotypes were clearly clustered into another group according to AFLP markers.

Glyphosate translocation in hairy fleabane (Conyza bonariensis) biotypes

The objective of this work was to evaluate the translocation of glyphosate in C. bonariensis plants resistant and susceptible to that herbicide. The 14C-glyphosate was mixed with commercial gyhphosate (800 g ha-1) and applied on the center of the adaxial face of a third node leaf, using a micro syringe, and adding 10 µL of a solution with specific activity of 1,400 Bq, 45 days after plant emergence. The concentration of the glyphosate translocated in the plant was evaluated at time intervals of 6, 12, 36 and 72 hours after being applied on the application leaf, stem, roots and leaves. Ten hours after treatment application, the distribution of the product in the application leaf, divided into base, center and apex, was also evaluated by measuring the radiation emitted by 14C-glyphosate in a liquid scintillation spectrometer. Greater glyphosate retention was observed in the resistant biotype leaf, approximately 90% of the total absorbed up to 72 hours. In the susceptible biotype, this value was close to 70% in the same period. Susceptible biotype leaves, stem and roots showed greater concentration of glyphosate, indicating greater translocation efficiency in this biotype. In the resistant biotype, the herbicide accumulated in greater quantity at the apex and center of the application leaf, while in the susceptible biotype greater accumulation was observed at the base and center leaf. Thus, it can be stated that the resistance mechanism is related to the differential translocation of this herbicide in the biotypes.

Effect of dose and application site on quinclorac absorption by barnyardgrass biotypes

This work aimed to evaluate the uptake and translocation of quinclorac in function of application sites (shoot or roots) by Echinochloa crusgalli biotypes resistant and susceptible to this herbicide. The treatments consisted of quinclorac doses (0; 0.5; 1; 2; 4; 16 and 64 ppm), applied on the shoot or roots of seedlings of barnyardgrass biotypes. The experimental units consisted of plastic cups containing 250 cm³ of sand. The treatments were applied 10 days after emergence, when barnyardgrass plants reached a 2- to 3- leaf growth stage. The barnyardgrass biotypes were irrigated with nutritive solution weekly and maintained for 40 days after emergence, when length, fresh and dry matter of shoot and roots were evaluated. Variance analysis was carried out using the F test at 5% probability, and in case of significance, a non-linear regression analysis was also carried out using a three-parameter logistic model. In the susceptible biotype, quinclorac was more absorbed by the roots than by the shoot. Comparing dry mass production of the different plant parts of the susceptible biotype per application site, it was verified that quinclorac action is higher when applied to the plant roots. However, for the resistant biotype, it was not possible to determine the dose causing 50% reduction in dry mass accumulation (GR50) and in the resistance index (RI) between both biotypes, due to its high resistance to quinclorac (128 times the recommended dosage). The results showed that quinclorac resistance by the evaluated biotype is not due to differences in the absorption site, strongly suggesting that the resistance acquired by the biotype may result from alteration in the target site.

Competitiveness of ALS inhibitors resistant and susceptible biotypes of Greater Beggarticks (Bidens subalternans)

The continuous use of ALS-inhibiting herbicides has led to the evolution of herbicide-resistant weeds worldwide. Greater beggarticks is one of the most troublesome weeds found in the soybean production system in Brazil. Recently, a greater beggarticks biotype that is resistant (R) to ALS inhibitors due to Trp574Leu mutation in the ALS gene was identified. Also, the adaptive traits between susceptible (S) and R to ALS inhibitors biotypes of greater beggarticks were compared. Specifically, we aimed to: (1) evaluate and compare the relative growth rates (RGR) between the biotypes; (2) analyze the seed germination characteristics of R and S biotypes under different temperature conditions; and (3) evaluate their competitive ability in a replacement series study. The experiments were conducted at the University of Arkansas, USA, in 2007 and at Universidade Federal do Rio Grande do Sul (Federal University of Rio Grande do Sul), Brazil, in 2008. Plant proportions for replacement series studies were respectively 100:0, 75:25, 50:50, 25:75 and 0:100, with a total population of 150 plants m-2. There was no difference in RGR between R and S biotypes. The R-biotype germination rate was lower than that of the S biotype. However, at low temperature conditions (15 ºC), the reverse was observed. In general, there is no difference in the competitive ability between R and S greater beggarticks biotypes.

Eletrolite leakage as a technique to diagnose euphorbia heterophylla biotypes resistant to ppo-inhibitors herbicides

The action of herbicides that affect the integrity of cell membranes and cause leakage, like PPO-inhibitors, can be detected by measuring the electric conductivity (EC) of a solution in which the plant tissue target is incubated in the presence of herbicide. The objectives of this work were to confirm PPO resistance in a new Euphorbia heterophylla (EPHHL) biotype, and to compare the electrolyte leakage from R and S to PPO-inhibitors biotypes, using two different methods of incubation in a solution containing herbicides. One experiment was carried in greenhouse and three in laboratory, with a completely randomized design. In the greenhouse experiment, four biotypes of EPHHL were sprayed with seven rates of fomesafen to confirm resistance in suspected biotypes. Leaf disks from R and S EPHHL biotypes in the second and the third experiments and entire leaves in the fourth experiment were incubated in a solution containing PPO-inhibitors to subsequently determine EC of solution. The study confirmed the resistance to PPO-inhibitors in two EPHHL biotypes. There were no significant differences between S and R biotypes in the experiments with the incubation of leaf disks, but incubation of entire leaves of EPHHL S biotype showed higher EC when in a solution with fomesafen, in comparison to the R biotype. The results of this work are an indirect evidence that resistance to PPO-inhibitors is related to lower absorption of herbicide by the shoots and also to some kind of mechanism to cope with oxidative stress.

Rapid diagnosis of resistance to imidazolinone herbicides in barnyardgrass (Echinochloa crus-galli) and control of resistant biotypes with alternative herbicides

The resistance of barnyardgrass (Echinochloa crus-galli) to imidazolinone herbicides is a worldwide problem in paddy fields. A rapid diagnosis is required for the selection of adequate prevention and control practices. The objectives of this study were to develop expedite bioassays to identify the resistance to imidazolinone herbicides in barnyardgrass and to evaluate the efficacy of alternative herbicides for the post-emergence control of resistant biotypes. Three experiments were conducted to develop methods for diagnosis of resistance to imazethapyr and imazapyr + imazapic in barnyardgrass at the seed, seedling and tiller stages, and to carry out a pot experiment to determine the efficacy of six herbicides applied at post-emergence in 13 biotypes of barnyardgrass resistant to imidazolinones. The seed soaking bioassay was not able to differentiate the resistant and susceptible biotypes. The resistance of barnyardgrass to imidazolinones was effectively discriminated in the seedlings and tiller bioassays seven days after incubation at the concentrations of 0.001 and 0.0001 mM, respectively, for both imazethapyr and imazapyr + imazapic. The biotypes identified as resistant to imidazolinones showed different patterns of susceptibility to penoxsulam, bispyribac-sodium and pyrazosulfuron-ethyl, and were all controlled with profoxydim and cyhalofop-butyl. The seedling and tiller bioassays are effective in the diagnosis of barnyardgrass resistance to imidazolinone herbicides, providing an on-season opportunity to identify the need to use alternative herbicides to be applied at post-emergence for the control of the resistant biotypes.

Germination and growth of Fimbristylis miliacea biotypes resistant and susceptible to acetolactate synthase-inhibiting herbicides

Weed biotypes resistant and susceptible to herbicides may have differences in their adaptive values. The aims of this study were to compare, under controlled and non-competitive condition, the growth analysis, germination features and seed weight of Fimbristylis miliacea (FIMMI) biotypes resistant and susceptible to acetolactate synthase (ALS) inhibiting herbicides. Experiments were conducted in a greenhouse and in a laboratory from October 2008 to February 2010. Two resistant biotypes (FIMMI 10 and FIMMI 12) and one susceptible biotype (FIMMI 13) were used for the studies. For the study on growth analysis, the treatments were arranged in a completely randomized experimental design with four replications and sampled at 21, 28, 35, 42, 49, 56, 69 days after emergence (DAE) and at flowering stage. For the studies on germination speed, germination and seed weight, the indexes for germination speed, percentage of germination at different temperatures and seed weight of the biotypes were determined. The results showed that the resistant biotype FIMMI 12 shows differences in all variables compared to the resistant biotype FIMMI 10 and compared to the susceptible biotype FIMMI 13, only for the evaluation at flowering. The susceptible biotype FIMMI 13 showed a higher germination speed index and higher germination rate when compared with the resistant biotypes. On the other hand, the resistant biotypes FIMMI 10 and FIMMI 12 showed higher seed weight.

Adaptive value of ryegrass biotypes with low-level resistance and susceptible to the herbicide fluazifop and competitive ability with the wheat culture

Ryegrass is the main weed in wheat crop, causing yield loss due to competition by environmental resources. The objectives of this study were to estimate the fitness cost of ryegrass biotypes with low-level resistance and susceptible to fluazifop and to investigate the relative competitive ability of these biotypes between themselves and against the crop. Thus, fitness cost and competitive ability experiments were conducted under greenhouse conditions. For the fitness cost experiments, the low-level resistant ryegrass biotypes and those susceptible to fluazifop were used. For competitive ability, the treatments were arranged in a replacement series, with five proportions of the wheat cultivar FUNDACEP Horizonte and the low-level resistant and susceptible ryegrass biotypes 100:0, 75:25, 50:50, 25:75 and 0:100. Competitive analysis was carried out through diagrams applied to the replacement experiments and use of relative competitiveness indices. Variables evaluated were plant height, in the fitness cost experiment, and leaf area and shoot dry biomass in both experiments. The ryegrass biotypes show overall similar fitness cost and competitive ability. The wheat cultivar FUNDACEP Horizonte is superior in competitive ability to the ryegrass biotype with low-level resistance and equivalent to the susceptible biotype.

Dose-response curves of Lolium multiflorum biotypes resistant and susceptible to clethodim

Chemical control with herbicides, especially glyphosate, is the main method used to control ryegrass. However, the repeated use of glyphosate has selected resistant ryegrass biotypes. Thus, the ACCase inhibitor herbicides have become the main alternative to control glyphosate-resistant biotypes, being widely used by farmers in Rio Grande do Sul. Repeated use of ACCase inhibitors, in turn, have selected ryegrass biotypes resistant to this herbicide mechanism. Thus, the objective of this study was to evaluate the response of ryegrass biotypes to different clethodim rates by dose-response curves. Increasing doses (0, 12, 24, 48, 72, 96, 144 and 192 g a.i. ha-1) of the herbicide clethodim were applied at the 3-4 ryegrass leaf stage. The variables control at 14 and 28 days after treatment (DAT) and shoot dry weight were evaluated. The data were fitted by nonlinear regression log-logistic and C50 and GR50 were calculated based on the equation. The resistance factor was obtained by the ratio of C50 or GR50 of the resistant biotype by matching the susceptible biotype. Based on the equation parameters, the doses of GR50 64.7 and 234.5 g a.i. ha-1 clethodim and C50 11.2 and 172.1 g a.i. ha-1 clethodim were obtained, at 28 DAT for the susceptible and resistant biotypes, respectively. The ryegrass biotype denominated Cotril is resistant to clethodim, being controlled with a dose 15.3 times greater than that of the susceptible biotype, and a 50% reduction of this biotype occurs with a dose 3.62 times higher than that of the susceptible one.

Glyphosate influence on the physiological parameters of Conyza bonariensis biotypes

The objective of this study was to determine changes in gas exchange and inhibition of EPSPs, based on the accumulation of shikimic acid in horseweed biotypes resistant and sensitive to glyphosate. Two experiments were conducted in a factorial model. The first one evaluated horseweed biotypes (one resistant and one susceptible to glyphosate), and herbicide rates (0 and 1,080 g a.e. ha ¹) applied on the weed. In the second experiment, the horseweed biotypes (susceptible and resistant to glyphosate) were evaluated in five periods as following: 0, 3, 7, 10, and 14 days after herbicide application (DAH). The photosynthetic rate, transpiration, carboxylation efficiency, and water efficiency were determined using an infrared gas analyzer (IRGA), and shikimic acid concentration by HPLC. The application of glyphosate damaged the photosynthetic parameters of the susceptible biotype, causing complete inhibition of the photosynthetic rate, transpiration rate, carboxylation efficiency and water use efficiency, starting from the 7 DAH. On the other hand, total inhibition of the photosynthetic parameters was not observed for the resistant biotype. Shikimic acid accumulation occurred in both biotypes after glyphosate application but the susceptible biotype had the highest concentrations, indicating greater sensitivity of the enzyme EPSPs. The accumulation of shikimic acid in the resistant biotype indicates that the mechanism of resistance is not related to the total insensitivity of the enzyme EPSPs to glyphosate and/or that other resistance mechanisms may be involved.

Determination of photosynthetic pigments in fleabane biotypes susceptible and resistant to the Herbicide Glyphosate

Chlorophylls and carotenoids are the main photosynthetic pigment in plants. In the weeds, the greatest amount of photosynthetic pigments can result in high competitiveness of the species. The aim of this study was to quantify the content of photosynthetic pigments in biotypes of fleabane (Conyza bonariensis) susceptible and resistant to glyphosate, by two different methods, as well as a correlation between chlorophyll content obtained by portable and classical methodology (extractable chlorophyll). An experiment was conducted in greenhouse and laboratory, 2 x 5 factorial scheme, where factor A was equivalent to biotypes of fleabane (resistant and susceptible to glyphosate) and factor B to developmental stages plants (rosette vegetative I, II and III and reproduction). At all stages of development, fleabane plants were evaluated with the portable determiner (chlorophyll content) and then the same leaves were subjected to classical methodology laboratory (extractable pigments). The resistant biotype of fleabane showed higher contents of chlorophyll a, b, and total carotenoids, inferring a greater competitive potential regarding the susceptible population to the herbicide. The portable determiner of chlorophyll showed high correlation with the classical method of determination of photosynthetic pigments, and can thus be used to accurately assess this, saving time and reagents.

Differential susceptibility of biotypes of conyza sumatrensis to Chlorimuron-ethyl herbicide

Horseweed (Conyza spp.) is an annual weed, infesting soybean crops in southern Brazil, with chlorimuron-ethyl being one of the most commonly used herbicides for its control. However, in recent soybean harvests, an unsatisfactory control of this weed using this herbicide was observed, generating suspicion regarding the selection of resistant biotypes. The objective of this study was to evaluate the susceptibility of horseweed biotypes to the herbicide chlorimuron-ethyl. Two experiments were conducted in a greenhouse; in the first one, the biotypes were selected selected, and the second experiment was arranged in a 5 x 5 factorial in a completely randomized design with four replications. The treatments used in the preparation of the dose response curves were doses of the herbicide chlorimuron-ethyl (0.0, 1.56, 3.13, 6.25, 12.5, and 25 g ha-1), applied on the five horseweed biotypes at the 3-4 leaf growth stage. The variables evaluated were visual control percentage and shoot dry weight, compared to the control without herbicide application, and plant acetolactate accumulation. It was concluded that there is a differential susceptibility among the biotypes at doses of less than 20 g ha-1 (dose response curves), which indicates low-level resistance. The practical consequences are the indications of chlorimuron-ethyl application at the maximum doses recomended and that the practice of rotating mechanisms of action must be used in the chemical weed management of these areas.

Shikimate accumulation, glyphosate absorption and translocation in horseweed biotypes

In Brazil, few research works on mechanisms of weed resistance to glyphosate have been conducted so far. Therefore, this research aimed to study analytical procedures determining the relation between the concentration of plant shikimate after glyphosate application and the plant resistance to this herbicide; and evaluate the glyphosate absorption and translocation into two resistant ® and susceptible (S) horseweed biotypes to glyphosate. Horseweed plants with nine true leaves received glyphosate (720 g a.e. ha-1), and 2, 3, 4, 7 and 10 days after application (DAA) the concentration of shikimic acid was measured by HPLC. In another experiment, plants were treated with radiolabeled glyphosate (14C) (1.456 MBq mmol-1 specific activity) and radioactivity was measured 4, 8, 24, 48 and 72 hours after treatment (HAT) by liquid scintillation spectrometry. The shikimate concentration in plants increased 16,351.14 and 7,892.25 mg kg-1 of dry weight, for R and S plants respectively, at seven DAA. Therefore, the procedure for quantification of shikimic acid was suitable for R and S plants differentiation to glyphosate, indicating that the R population is actually resistant to glyphosate. On average, 98% of glyphosate applied was absorbed by the studied biotypes, at 72 HAT. Around 68% of the absorbed radioactivity remained on the biotypes leaves treated, the S biotype showing the highest translocation. Therefore, the R biotype resistance mechanism studied is associated to the differential translocation.

MULTIPLE RESISTANCE OF Sagittaria montevidensis BIOTYPES TO ACETOLACTATE SYNTHASE AND PHOTOSYSTEM II INHIBITING HERBICIDES

ABSTRACT The objective of this research was to evaluate the occurance of multiple resistance of Sagittaria montevidensis (SAGMO) biotypes to acetolactate synthase (ALS) and photosystem II (PSII) inhibiting herbicides through dose-response experiments. The experiment was conducted in a greenhouse from October 2012 to March 2013, in Pelotas, RS. The experimental design was completely randomized, with four replications. Treatments were arranged in a triple factorial design: two biotypes of S. montevidensis(SAGMO 35 - susceptible to herbicides and SAGMO 32 - suspected to be multiple resistance to ALS and PSII inhibiting herbicides), four herbicides (penoxsulam, (imazethapyr+imazapic), bentazon and saflufenacil) and 8 rates of these herbicides (1/32x, 1/16x, 1/8x, 1/4x, 1/2x, 0x, 1x, 2x, 4x, 8x, 16x, 32x and 64x). SAGMO 32 biotype presented high levels of resistance to penoxsulam, (imazethapyr+imazapic) and bentazon. For a 50% reduction in dry matter of the resistant biotype rate of 138 and 2.46 times higher than the label required for the susceptible biotype of the herbicides (imazethapyr+imazapic) and bentazon, respectively, are required. Saflufenacil may be used successfully to controlSagittaria montevidensis resistant in irrigated rice.