Evaluation of the antimicrobial activity of Eucalyptus radiata essential oil against Escherichia coli strains isolated from meat products.

The present study sought to evaluate the antimicrobial and anti-adherent potential of Eucalyptus radiata essential oil against food-borne strains of Escherichia coli. The study was performed using the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). In addition, the disk diffusion technique was used to evaluate the association of Eucalyptus radiata essential oil with synthetic antimicrobials. The Minimum Inhibitory Adherence Concentration (MIC) was also performed. The results revealed that E. radiata showed antimicrobial activity against the E. coli strains tested, with MIC values ranging from 500 µg/mL to 1000 µg/mL and MBC values ranging from 500 µg/mL to 1,024 µg/mL. As for the associations, it was observed that E. radiata oil exhibited a synergistic effect for some antibiotics, especially Ceftriaxone, with greater interference from the essential oil. Furthermore, it was effective in inhibiting the adherence of bacterial strains of E. coli, showing a more significant antibiofilm effect than the antibacterial agent 0.12% chlorhexidine digluconate. In summary, the essential oil of E. radiata showed antimicrobial potential against strains of E. coli of food origin, and can therefore, through in-depth studies, be used alone or in association with synthetic antimicrobials to combat infections caused by this pathogen.

Evaluation of the antibacterial activity of trans-anethole against Enterococcus cloacae and Enterococcus faecalis strains of food origin.

The present study sought to evaluate the antibacterial activity of trans-anethole against food-borne strains of Enterobacter cloacae and Enterococcus faecalis. The study was performed using Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC) methods, in addition, disc diffusion technique was used to evaluate the association of trans-anethole with synthetic antimicrobials. Minimum Inhibitory Concentration for Adherence (MICA) testing was also performed. The results revealed that trans-anethole presents no antibacterial activity at any of the concentrations used against the E. cloacae strains tested. However, trans-anethole presented antibacterial effect against five of the six E. faecalis bacterial strains tested, with MIC values ranging from 500 µg/mL to 1000 µg/mL. Further, when analyzing the MBC results against E. faecalis, it was observed that the compound presented values ranging from 500 µg/mL to 1000 µg/mL. As for the associations, it was observed that trans-anethole when combined with the antimicrobials ampicillin, gentamicin, ciprofloxacin, and ceftriaxone presented synergistic effect against most strains of E. faecalis. However, both trans-anethole and the control chlorhexidine (0.12%) presented no antibiofilm effects against strains of E. faecalis. In short, trans-anethole presented potential antibacterial against E. faecalis strains of food origin, and may upon further study, it may be used alone or in association with synthetic antimicrobials to combat infections caused by this bacterium.

Novel MLST sequence types of pathogenic Leptospira spp.: Opening the black box of animal leptospirosis in Brazil.

In the current context of the emergence of certain infectious diseases and discussion of the One Health concept for many of these, the study of leptospirosis - both in domestic and wild hosts - cannot be neglected. The study of animal leptospirosis has evolved in recent years. It has been demonstrated that the human-animal-environment interface is more important than previously thought. In the present study, 35 strains of five pathogenic Leptospira species were isolated from different animal species in Brazil and characterized by rrs, secY, and Multilocus Sequence Typing (MLST) sequencing. Phylogenetic inferences were performed and the molecular diversity of the populations (intra- and inter-population levels) was evaluated. Among the five studied species, 18 different sequence types (STs) were found (22 new alleles and 11 new STs). eBURST analysis revealed two clonal complexes (CCs) and seven singletons. A high genetic diversity was demonstrated (H = 0.954 ± 0.017), mainly for the L. santarosai population (H = 0.942 ± 0.034, n = 20). The same strain was identified in different host species, as well as strains with zoonotic potential circulating in the country. Although the difficulty of culturing Leptospira strains is well known, the high variability of the strains found in Brazil highlights the importance of animals in maintaining the biological cycle of the bacterium in nature. Moreover, the selection of autochthonous strains for the development of vaccines becomes a challenge.

Detection of East/Central/South African genotype Chikungunya virus during an outbreak in a southeastern state of Brazil.

Chikungunya virus (CHIKV) is an arbovirus transmitted by Aedes mosquitoes that was first identified in Brazil in 2014. It causes a febrile illness characterised by severe arthralgia and rash. Our group investigated a suspected CHIKV outbreak in Governador Valadares, state of Minas Gerais, Brazil and from 25 acute-phase patients, 10 had qRT-PCR positive sera samples and had E1 partial sequence amplified and Sanger sequenced. Samples were identified as East/Central/South African (ECSA) genotype by phylogenetic analysis and clustered with CHIKV sequences isolated in the neighbour state of Bahia. Our findings confirm previous predictions that ECSA genotype would spread through northeast and southeast of Brazil.

High frequency of leptospiral vaginal carriers among slaughtered cows.

Bovine leptospirosis is one of the most important reproductive diseases that compromise the productivity of cattle farming. However, the presence of the agent on vaginal environment is still poorly understood in cattle. Considering this context, the present study aimed to detect the presence of pathogenic Leptospira sp. in vaginal fluid (VF) of cows. VF and urine were collected from 254 cows from a slaughterhouse for bacteriological culture and PCR (lipL32 gene). Overall, eleven pure culture (4.3%) of leptospiral isolates were obtained. Leptospiral DNA was detected in 128 (50.4%) of VF samples and 81 (31.0%) of urine samples, while on 75 (29.5%) it was exclusively in VF and 28 (11.3%) only in the urine. Detection of leptospiral DNA and the recovery of viable leptospires from VF of a high number of cows without apparent symptoms highlight the role of vaginal carriers and indicate that venereal transmission (female-to-male) could occur in that species. Moreover, VF should be encouraged as a valuable sample for diagnosis of bovine genital leptospirosis.

Molecular Characterization and Serology of Leptospira kirschneri (Serogroup Grippotyphosa) Isolated from Urine of a Mare Post-Abortion in Brazil.

A strain of Leptospira kirschneri (serogroup Grippotyphosa) was cultured from urine of a mare post-abortion in Brazil and characterized by serogrouping, multiple-locus variable-number tandem repeat analysis, PGFE, and sequencing of genes rrs and secY. Strains of L. kirschneri have apparently never been recovered from horses in tropical area, only in Europe and USA. Knowledge of local epidemiology is important to interpret genetic profiles of leptospires circulating in an area.

Genotyping of Leptospira directly in urine samples of cattle demonstrates a diversity of species and strains in Brazil.

The aim of this study was to identify Leptospira in urine samples of cattle by direct sequencing of the secY gene. The validity of this approach was assessed using ten Leptospira strains obtained from cattle in Brazil and 77 DNA samples previously extracted from cattle urine, that were positive by PCR for the genus-specific lipL32 gene of Leptospira. Direct sequencing identified 24 (31·1%) interpretable secY sequences and these were identical to those obtained from direct DNA sequencing of the urine samples from which they were recovered. Phylogenetic analyses identified four species: L. interrogans, L. borgpetersenii, L. noguchii, and L. santarosai with the most prevalent genotypes being associated with L. borgpetersenii. While direct sequencing cannot, as yet, replace culturing of leptospires, it is a valid additional tool for epidemiological studies. An unexpected finding from this study was the genetic diversity of Leptospira infecting Brazilian cattle.

Is glycated hemoglobin related to other dysmetabolic variables implicated in the increase of cardiovascular risk in polycystic ovary syndrome? A comparative study.

BACKGROUND: In non-PCOS patients the concentration of glycated hemoglobin (HbA1C) has been employed to identify individuals at higher risk for impaired glucose tolerance (IGT) and diabetes mellitus. A few studies have examined the role of HbA1C in PCOS patients and current results are controversial. AIM: To compare the strength of the association between glycated hemoglobin and other predictors of cardiovascular risk in polycystic ovary syndrome (PCOS). METHODS: This cross-sectional study enrolled 197 PCOS patients and 72 non-PCOS women. Transvaginal ultrasound, biochemical and hormone measurement were performed. Glycated hemoglobin (HbA1C) was correlated with other variables related to dysmetabolic/vascular diseases. RESULTS: The HbA1C levels were 6.0±1.4% and 4.9±0.4% in PCOS patients and non-PCOS controls, respectively (p<0.001). The HbA1C levels were≥5.7% in 46.4% of PCOS and in none of the control subjects (OR=90.8). HbA1C was well-correlated with several anthropometric, metabolic and endocrine parameters. Stepwise multiple regression including HbA1C and other known predictors of cardiovascular risk resulted in a significant model in which body mass index (BMI) and free testosterone exhibited the best correlation with HbA1C (adjusted R(2)=0.530; F=39.8; p<0.001). CONCLUSION: HbA1C was elevated and correlated with anthropometric, biochemical and endocrine variables of metabolic/vascular disease risks in PCOS patients. Combined HbA1C, BMI and free testosterone levels provided a significant model with potential use to evaluate metabolic/vascular disease in PCOS patients.

Isolation of Leptospira interrogans Hardjoprajitno from vaginal fluid of a clinically healthy ewe suggests potential for venereal transmission.

A total of 15 adult ewes from one flock known to be seroreactive for leptospirosis was studied. Urine and vaginal fluid were collected from each animal to test for the presence of leptospires using bacterial culture and conventional PCR methods. One pure culture of Leptospira sp. was obtained from the vaginal fluid sample of a non-pregnant ewe. The isolate was characterized by DNA sequencing of the rrs and secY genes, variable-number of tandem-repeats (VNTR) analysis and serogrouping, and the isolate was typed as Leptospira interrogans serogroup Sejroe serovar Hardjo type Hardjoprajitno. This report indicates the presence of viable Leptospira in the vaginal fluid of a ewe, suggesting the potential for venereal transmission of leptospires in sheep.

Potential differences between Leptospira serovars, host-adapted (Bratislava) and incidental (Copenhageni), in determining reproductive disorders in embryo transfer recipient mares in Brazil.

The objective was to investigate potential differences between two Leptospira serovars, host-adapted (Bratislava) and incidental (Copenhageni), in causing reproductive disorders in mares. From August 2009 to March 2011, 608 adult recipient mares from eight studs located in the state of Rio de Janeiro, Brazil, were screened for leptospirosis. These mares were 3-8 years of age, of various breeds, and were managed in a semiextensive system (embryo transfer centres). According to the reproductive history of these mares, the studs were categorised as Group A (357 mares: high prevalence of reproductive problems), and Group B (251 mares: all pregnant). Of the 608 samples tested serologically, 273 (44.9 per cent) were reactive (titres ≥200), predominantly against Bratislava (62.3 per cent of reactive mares). In Group A, 247/357 sera (69.2 per cent) were reactive, with reactivity against Bratislava (64.8 per cent of reactive). By contrast, in Group B, only 26/251 sera (10.4 per cent) were reactive, the majority against Copenhageni (61.5 per cent of reactive). Seroreactivity against Bratislava was more likely to be associated (P<0.001) with reproductive problems than seroreactivity against Copenhageni; this association was true for early embryonic death (P<0.001), perinatal death (P<0.01), and abortion (P<0.001). Additionally, 29 urine samples were collected (from Group A mares) for Leptospira species PCR, of which 16 were positive (55.2 per cent). We concluded that leptospirosis was associated with poor reproductive performance in embryo recipients, with the majority due to host-adapted Bratislava.

Urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.

The aim of the present study was to consider the wide usage of urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. A total of 512 adult animals (300 cattle, 138 horses, 59 goats and 15 pigs), from herds/flocks with reproductive problems in Rio de Janeiro, Brazil was studied by serology and urinary PCR. From the 512 serum samples tested, 223 (43.5 %) were seroreactive (cattle: 45.6 %, horses: 41.3 %, goats: 34%and pigs: 60 %). PCR detected leptospiral DNA in 32.4 % (cattle: 21.6 %, horses: 36.2 %, goats: 77.4 % and pigs: 33.3 %. To our knowledge there is no another study including such a large number of samples (512) from different species, providing a comprehensive analysis of the usage of PCR for detecting leptospiral carriers in livestock. Serological and molecular results were discrepant, regardless the titre, what was an expected outcome. Nevertheless, it is impossible to establish agreement between these tests, since the two methodologies are conducted on different samples (MAT - serum; PCR - urine). Additionally, the MAT is an indirect method and PCR is a direct one. In conclusion, we have demonstrated that urinary PCR should be considered and encouraged as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.

Nutritional variables and work-related accidents: a case-control study.

BACKGROUND: Nutritional aspects are important for the prevention of diseases and disorders, and few studies have focused on the relationship between risk of work injury and nutritional variables. OBJECTIVE: This study aimed to verify whether nutritional variables constitute risk factors for work-related accidents. PARTICIPANTS: 1,422 industrial workers (600 cases plus 822 controls). METHODS: A case-control study was carried out in an industrial city in south-east Brazil. A multiple logistic regression model was adjusted using work-related accidents as the response variable and nutritional variables as predictors. The associations were assessed by Odds Ratio (OR), with a p-value < 0.05. RESULTS: 47.29% of the workers were overweight or obese. Protective factors for work-related accidents were (a) attending formal education for an above average number of years (OR=0.91, p< 0.0001) and (b) eating a traditional dinner (OR=0.67, p=0.0087). Risks factors were (a) hard physical effort in the workplace (OR=1.37, p< 0.0001), (b) having lunch in the workplace (OR=1.57, p<0.0001) and (c) receiving government benefits in the form of food stamps (OR=1.39, p=0.0350) or food baskets (OR=1.30, p=0.0414). CONCLUSION: Our findings suggest an association between nutritional variables and work-related accidents. This indicates the need, during the formulation of policies for these kinds of government benefits, to include nutrition aspects in order to minimize work-related accidents risks.

The role of horses in the transmission of leptospirosis in an urban tropical area.

The objective of this study was to demonstrate the presence of leptospires in equine urine, as evidence for a potential role of horses in transmission of this organism. Thoroughbred horses (aged 2-5 years, n = 276) from Rio de Janeiro, Brazil, were studied. After a severe storm, the premises of the animals remained flooded for 72 h. Blood samples for serology were collected on days 20 and 35 (day of storm = day 0). On day 20, 132 (47·8%) horses were seroreactive (titre ≥200) and, of these, 23 (31·0%) had increased antibody titres on day 35. Furthermore, 34 urine samples (for PCR and culture) were collected from seroreactive horses on day 35. Copenhageni was the most frequent serovar (88·8% of reactive titres). Although none of the urine samples were culture positive, 12 (35·2%) were PCR positive. This is apparently the first report of evidence of leptospires in urban horses. Furthermore, we suggest that these animals can play a role in the transmission of leptospirosis in urban areas.

High-frequency electroacupuncture versus carprofen in an incisional pain model in rats

The objective of the present study was to compare the effect of electroacupuncture (EA) and carprofen (CP) on postoperative incisional pain using the plantar incision (PI) model in rats. A 1-cm longitudinal incision was made through skin, fascia and muscles of a hind paw of male Wistar rats and the development of mechanical and thermal hypersensitivity was determined over 4 days using the von Frey and Hargreaves methods, respectively. Based on the experimental treatments received on the third postoperative day, the animals were divided into the following groups: PI+CP (CP, 2 mg/kg, po); PI+EAST36 (100-Hz EA applied bilaterally at the Zusanli point (ST36)); PI+EANP (EA applied to a non-acupoint region); PI+IMMO (immobilization only); PI (vehicle). In the von Frey test, the PI+EAST36 group had higher withdrawal force thresholds in response to mechanical stimuli than the PI, PI+IMMO and PI+EANP groups at several times studied. Furthermore, the PI+EAST36 group showed paw withdrawal thresholds in response to mechanical stimuli that were similar to those of the PI+CP group. In the Hargreaves test, all groups had latencies higher than those observed with PI. The PI+EAST36 group was similar to the PI+IMMO, PI+EANP and PI+CP groups. We conclude that 100-Hz EA at the ST36 point, but not at non-acupoints, can reduce mechanical nociception in the rat model of incisional pain, and its effectiveness is comparable to that of carprofen.

High-frequency electroacupuncture versus carprofen in an incisional pain model in rats.

The objective of the present study was to compare the effect of electroacupuncture (EA) and carprofen (CP) on postoperative incisional pain using the plantar incision (PI) model in rats. A 1-cm longitudinal incision was made through skin, fascia and muscles of a hind paw of male Wistar rats and the development of mechanical and thermal hypersensitivity was determined over 4 days using the von Frey and Hargreaves methods, respectively. Based on the experimental treatments received on the third postoperative day, the animals were divided into the following groups: PI+CP (CP, 2 mg/kg, po); PI+EAST36 (100-Hz EA applied bilaterally at the Zusanli point (ST36)); PI+EANP (EA applied to a non-acupoint region); PI+IMMO (immobilization only); PI (vehicle). In the von Frey test, the PI+EAST36 group had higher withdrawal force thresholds in response to mechanical stimuli than the PI, PI+IMMO and PI+EANP groups at several times studied. Furthermore, the PI+EAST36 group showed paw withdrawal thresholds in response to mechanical stimuli that were similar to those of the PI+CP group. In the Hargreaves test, all groups had latencies higher than those observed with PI. The PI+EAST36 group was similar to the PI+IMMO, PI+EANP and PI+CP groups. We conclude that 100-Hz EA at the ST36 point, but not at non-acupoints, can reduce mechanical nociception in the rat model of incisional pain, and its effectiveness is comparable to that of carprofen.

Conservação de inimigos naturais (Insecta) em tomateiro orgânico / Conservation of natural enemies (Insecta) in the organic tomato crop

A adoção de práticas culturais como o consórcio de culturas e o tipo de irrigação podem beneficiar a comunidade de inimigos naturais no agroecossistema ao disponibilizar micro-habitas mais favoráveis e recursos alternativos, principalmente em períodos de baixa precipitação pluviométrica. O objetivo deste trabalho foi avaliar como o consórcio tomate + coentro e o tipo de irrigação (gotejamento e aspersão) podem beneficiar a comunidade de inimigos naturais no agroecossistema do tomateiro. O trabalho foi realizado no campo experimental da Embrapa Hortaliças, Gama, DF, de setembro a novembro de 2008. Os tomateiros foram plantados em monocultura ou consorciados com o coentro e irrigados por gotejamento e por aspersão (três repetições por tratamento), formando dois conjuntos de parcelas experimentais de acordo com o tipo de irrigação. A comunidade de inimigos naturais foi amostrada por observações diretas em 20 plantas de tomate por parcela e nas plantas de coentro sacudindo-se as plantas em cima de uma bandeja onde eram coletados os insetos. A abundância, riqueza e diversidade das espécies de inimigos naturais foram maiores nas parcelas cultivadas com coentro, independente do sistema de irrigação. Nas parcelas plantadas em monocultura foram encontradas mais espécies quando o tomateiro foi irrigado por aspersão. No entanto, a resposta de cada espécie de inimigo natural ou grupo de espécies foi distinta para o consórcio ou o tipo de irrigação. Portanto, em períodos de baixa precipitação, o consórcio tomate + coentro e a irrigação por aspersão podem favorecer a conservação de inimigos naturais no agroecossistema do tomateiro orgânico.

The adoption of cultural practices such as companion plants and the kind of irrigation system can benefit the community of natural enemies in the agroecosystem due to the increase of more favorable microhabitats available and as a source of alternative resources, mainly in periods of low precipitation. This study evaluated how the tomato + coriander intercrop and the kind of irrigation (drip and sprinkler) could benefit the community of natural enemies in the tomato agroecosystem. This work was carried out in the experimental field of Embrapa Hortaliças, Gama, DF, from September to November 2008. The tomatoes were planted in monoculture or with coriander (companion plant) and irrigated by drip and sprinkler irrigation (three replicates per treatment), forming two groups of experimental plots according to the kind of irrigation. The community of natural enemies was sampled by the direct observation of specimens on 20 tomatoes plants per plot and in the treatments. Coriander plants were shaken over a plastic tray for collection of insects. The abundance, richness and diversity of natural enemies were higher in tomato + coriander plots, regardless of the irrigation system. We observed more species of natural enemies in tomato monoculture plots when it was irrigated by sprinkler irrigation. However, the cultural practices adopted had a different effect on each species or group of species. Therefore, in periods of low precipitation, the tomato + coriander consortium associated with sprinkler irrigation can enhance the conservation of natural enemies in the organic tomato agroecosystem.