Occurrence of Campylobacter jejuni and C. coli on broiler carcasses after chilling in southern Brazil

Campylobacter jejuniand C. colihave been associated with gastrointestinal disorders in human beings, due mainly to the consumption of chicken meat. Despite control measures for reducing contamination by these bacteria, the detection of Campylobacter in carcasses after chilling remains high.A total of 105 carcasses were assessed by the horizontal detection method in five federally inspected slaughterhouses in southern Brazil in 2012 and in the first three months of 2013. Campylobacterwas isolated in 37.1% of the carcasses, of which 97.5% contained C. jejuni and 2.5% were infected by C. coli. The rate of positive carcasses across the slaughterhouses ranged from 0 to 71.4%. Determining the occurrence of Campylobacteramong flocks is crucial for estimating the microbial load at specific points along the slaughtering process and for minimizing the risk of contamination of end products by Campylobacter.

Occurrence and characterization of Campylobacter spp.isolates in dogs, cats and children

To improve the understanding of implications of Campylobacterspp. infections in pets and children of different environments were analysed 160 faecal samples from children and 120 from pets (103 dogs and 17 cats). Campylobacter spp. were detected in 6.87% of the children and in 18.3% of the dogs and cats. From 33 stool samples positive for Campylobacter spp., 57.6% were identified as C. jejuni, and 33.4% were identified as C. coli. More than 50% of the isolates in pets were resistant to ceftiofur, sulphazotrim, norfloxacin and tetracycline. In humans, most of the isolates were resistant to amoxicillin, cefazolin, ceftiofur, erythromycin and norfloxacin. From 19 isolates of C. jejuni, 11 isolates from children and 5 from dogs contained two to four of the virulence genes flaA, pldA, cadF or ciaB.We found an association between the presence of virulence genes and diarrhoea. Furthermore, an association was observed between the presence of Campylobacter spp. and diarrhoea in dewormed pets with blood picture suggestive of bacterial infection, and the therapeutic use of antibiotics was associated with more positive detection of Campylobacterspp. in the faeces of pets. Our data indicate that virulent strains of Campylobacter spp. can be risk factor to diarrhoea in animals, and that high resistance to antimicrobial agents is common in pets.

Detecção de resistência às fluoroquinolonas em Campylobacter isolados de frangos de criação orgânica

Resumo Estudos têm revelado que a resistência às quinolonas em cepas de Campylobacter está relacionada à presença da mutação Treonina-86 para Isoleucina. Com o objetivo de investigar a presença dessa mutação em cepas de Campylobacter sensíveis e resistentes à ciprofloxacina e enrofloxacina, o conteúdo cecal de 80 frangos de corte de criação orgânica, abatidos sob Serviço de Inspeção Estadual (S.I.E.) do Estado do Rio de Janeiro, foram coletados e investigados para a presença de Campylobacter. A determinação da resistência à ciprofloxacina e enrofloxacina foi feita pela técnica de difusão em disco e de diluição em ágar para determinação da Concentração Inibitória Mínima (CIM). A detecção da mutação na Região Determinante de Resistencia às Quinolonas (RDRQ) no gene gyrA foi realizada através de sequenciamento. Campylobacter foi isolado a partir de 100% das amostras avaliadas, sendo 68,75% correspondente à C. jejuni e 31,25% à C. coli. No teste de difusão em disco, 100% das cepas foram resistentes à ciprofloxacina e 56,25% das cepas foram resistentes à enrofloxacina. No teste de diluição em ágar, todas as cepas foram resistentes à ciprofloxacina apresentando CIM variando de ≥ 16-64μg/mL, e resistência ou resistência intermediaria à enrofloxacina foi detectada em 42,50% (CIM ≥ 4-32μg/mL) e 38,75% (CIM = 2μg/mL) das cepas, respectivamente. A mutação Tre-86-Ile, foi observada em 100% das cepas analisadas. Além dessa mutação, foram observadas outras mutações não silenciosas (Val-73-Glu, Ser-114-Leu, Val-88-Asp, Ala-75-Asp, Ser-119-Gli, Arg-79-Lis) e mutações silenciosas (His-81-His, Ser-119-Ser, Ala-120-Ala, Fen-99-Fen, Ala-122-Ala, Gli-74-Gli, Ile-77-Ile, Ala-91-Ala, Leu-92-Leu, Val-93-Val, Ile-106-Ile, Tre-107-Tre, Gli-113-Gli, Ile-115-Ile, Gli-110-Gli). A observação de que cepas sensíveis à enrofloxacina pelos testes fenotípicos apresentavam a substituição Tre-86 para Ile sugere que outros mecanismos podem contribuir para a resistência à enrofloxacina em Campylobacter.

Resistance to β-lactam and tetracycline in Campylobacter spp.isolated from broiler slaughterhouses in southern Brazil

Abstract The study was carried out to screen and analyze the genetic characteristics of antimicrobial resistance in Campylobacter spp. from poultry sources. A total of 141 strains of Campylobacter isolated from samples of broilers of slaughterhouses in southern Brazil was identified by phenotypic and genotypic methods. Campylobacter isolates were evaluated for its antimicrobial susceptibility and the presence of resistance genes. The strains were investigated for antimicrobial susceptibility against two agents (ampicillin and tetracycline) by disk diffusion method. PCR assay was used to confirm the specie and the presence of ampicillin (blaOXA-61), tetracycline tet(O), and the energy-dependent multi-drug efflux pump (cmeB) genes. Campylobacter jejuni was the most ubiquitous; its presence was determined in 140 samples out of 141 (99.3%), whereas Campylobacter coli was found only in one of the contaminated samples (0.70%). The results obtained showed 65% and 35.5% of Campylobacter isolates resistant to β-lactams and tetracyclines, respectively. The cmeB gene responsible for multidrug resistance was detected in 26 isolates out 141 strains (18.5%). Moreover, 36 out of 141 Campylobacter strains (25.6%) were found to be resistant to at least two different antimicrobia resistance markers (β-lactams and tetracyclines).

Detecção dos genes codificantes da toxina CDT, e pesquisa de fatores que influenciam na produção de hemolisinas em amostras de Campylobacter jejuni de origem avícola

Resumo: Membros termofílicos do gênero Campylobacter são reconhecidos como importantes enteropatógenos para o ser humano e animais. A grande diversidade ecológica destes micro-organismos em diferentes habitats tais como água, animais e alimentos predispõem ao aparecimento de novos fatores de virulência. Este trabalho teve por objetivo detectar os genes codificantes da Toxina Distensiva Citoletal (CDT) por meio da técnica de PCR, pesquisar a atividade de hemolisinas e a influência de soluções quelantes e de íons nesta atividade. Foram utilizadas 45 amostras de Campylobacter jejuni de origem avícola para pesquisa de atividade hemolítica, cultivadas em Caldo Triptona de Soja (TSB). Após o crescimento bacteriano, as amostras foram semeadas em Ágar tríptico de soja (TSA) contendo 5% de sangue de ovino. Para verificar a influência de agentes quelantes e solução de íons na atividade hemolítica, as amostras de C. jejuni foram cultivadas em TSB contendo separadamente os quelantes EDTA, ácido acético, soluções de íons CaCl2, MgCl2 e FeCl3, em atmosfera de microaerofilia. Quanto à atividade de hemolisina de C. jejuni em placas de TSA - sangue ovino foi possível observar que houve hemólise em 40% das amostras analisadas apenas com caldo TSB. Somente o ácido acético apresentou ação quelante sobre a atividade de hemolisinas em amostras de C. jejuni semeadas em placas de TSA - sangue ovino. Para detecção dos genes cdtA, cdtB e cdtC através da técnica da Reação em Cadeia da Polimerase (PCR) foram utilizadas 119 amostras de C. jejuni de origem avícola. Foi possível observar que 37,8% possuíam o perfil de genes cdtABC. Os resultados demonstraram em amostras avícolas a presença de cepas de C. jejuni com potencial virulento, devido à presença dos genes da toxina CDT e potencial hemolítico, que apresentou ação reduzida in vitro com ácido acético.

Response of Grilage-Like Structures by the Mobility Approach

This work presents an application of the Mobility Approach to the analysis of the power flow through grillage-like structures. Such structures are usually found in offshore platforms, supporting large and heavy machines. Different wave kinds (longitudinal, flexural and torsional) were initially considered in the power flow analysis between two beams joined in L. Beams excited by an in-plane point force showed strong coupling between longitudinal-flexural waves, while that for out-of-plane point force excitation, flexural-torsional waves coupling represents the most important mechanism of energy transmission. The response determination of grillages by the mobility approach requires the structure to be separated into simple beam-like structural components. Equations for rotations and displacements at the joints of all beams are written for as mobility functions, and moments and forces acting at the joints. A system of equations relating all such internal forces and moments is obtained. This approach was applied to simple grillages. Response results showed good agreement when compared to those provided by Finite Elements.

The Environmental Status of the Port of HaminaKotka

This study is a part of the Ecologically Friendly Port Ust-Luga (EFP) project. The purpose of this study is to examine the environmental status of the Finnish ports and, more specifically, the Port of HaminaKotka. An analysis of the environmental status is performed mainly as a literature review, because the Finnish ports must comply with Finnish and EU legislation and with the binding international regulations and conventions created by different organizations. The International Maritime Organisation (IMO) has done groundbreaking work in the field of maritime safety and maritime environmental protection. The MARPOL convention has a great impact on decreasing pollution from international shipping and it applies to 99% of the world’s merchant tonnage. Pollution prevention covers: Oil pollution, Chemical pollution, Air pollution and GHG Emissions, Dumping of Wasted and Other Matters, Garbage, Sewage, Port Reception Facilities, Special Areas under MARPOL and Particularly Sensitive Sea Areas. There is also Pollution Prevention for other treaties like anti-fouling systems used on ships, the transfer of alien species by ships’ ballast water and the environmentally sound recycling of ships. There are more than twenty different EU and international regulations that influence ports and port operations in Finland. In addition, there is also national legislation that has an effect on Finnish ports. For the most part, the legislation for ports is common in the EU area, but the biggest and most important difference between the legislation in Finland and other EU countries is due to the Act on Environmental Impact Assessment Procedure. The Act states that the environmental impact assessment procedure shall be applied to projects that may have significant adverse environmental impacts, due to the special features of Finland`s nature and environment. In this Act, the term environmental impact refers to the direct and indirect effects inside and outside Finnish territory of a project or operations on human health, living conditions and amenity; soil, water, air, climate, organisms, interaction between them and biodiversity; community structure, buildings, landscape, townscape and cultural heritage; utilization of natural resources. In Finland, the Environmental Permit requires that ports collect all necessary information concerning environmental effects and make required reports to the Finnish authorities, stakeholders and the public. Commonly, environmental reporting is public and environmental achievements are emphasized in reporting and in media. At the moment, the problem in environmental reporting is that it’s difficult to compare data from different ports. There is enough data concerning the environmental effects and performance, but the manner of reporting and the quality of the data varies between ports. There are differences in the units and codes used, in some cases the information is not sufficient and it can even be rather unreliable. There are also differences regarding the subjects that are emphasized in reporting.

Soil organisms associated to the weed suppressant Crotalaria juncea (fabaceae) and its importance as a refuge for natural enemies

Soil organisms play an important role in organic crops of Crotalaria juncea (Fabaceae) and are associated with the natural conservation of the environment. The present study was aimed to investigate the population of soil organisms in the organic culture of C. juncea, as well as its importance as a refuge for natural enemies. Dalbulus maidis (Hemiptera: Cicadellidae), Diabrotica sp. (Coleoptera: Chrysomelidae), Doru luteipes (Dermaptera: Forficulidae), Gryllus assimilis (Orthoptera: Gryllidae), Lagria villosa (Coleoptera: Lagriidae), Melanotus sp. (Coleoptera: Elateridae), Meloidogyne incognita (Tylenchida: Heteroderidae), Nephila clavipes (Araneae: Nephilidae), Orius insidiosus (Hemiptera: Anthocoridae), Pheidole sp. (Hymenoptera: Myrmicidae), Phyllophaga sp. (Coleoptera: Scarabeidae), Procornitermes sp. (Isoptera: Termitidae), Solenopsis sp. (Hymenoptera: Formicidae), and Utetheisa ornatrix (Lepidoptera: Arctiidae) were identified in C. juncea. The organisms that were found during a 3-month period in 144 trenches in C. juncea were pest species (84.47%) and natural enemies (15.53%) as well. Natural enemies had an average of 11.89 individuals per 1.08 m³ of soil cultivated with C. juncea. The abundance of organisms in the pod stage (5.49%) of C. juncea was lower than that in the vegetative (83.50%) and flowering (11.01%) stages. Crotalaria juncea plants can be used as part of a crop system for Integrated Pest Management.

Imazapyr herbicide efficacy on floating macrophyte control and ecotoxicology for non-target organisms

the aims of this study were to determine imazapyr efficacy for floating macrophyte control and ecotoxicology for non-target organisms. For the floating macrophyte control efficacy tests were used the doses of 0,5; 1,0; 2,0; 2,5; 3,0; 3,5 and 4,0 L ha-1 and a control with 10 replicates. The acute toxicology for non-target organisms was estimated by lethal concentration 50% (LC50 and EC50). The floating macrophyte control efficacy was over 90%. Imazapyr was classified as moderately toxic for the following biomarkers: L. minor, H. eques, B. rerio, P. caudimaculatus, P. canaliculata, and P. mesopotamicus and lightly toxic for A. caroliniana. Thus, imazapyr herbicide is a tool with great potential to be used on floating macrophyte control (E. crassipes, P. stratiotes e S. molesta) in Brazil and this practice can be evaluated by the use of application biomarkers.

Cyanobacteria from coastal lagoons of Southern Brazil: coccoid organisms

Considering the great ecological importance of the cyanobacteria and the need for more detailed information about these organisms in Brazilian waters, this paper provides taxonomic information about the unicellular cyanobacteria flora in lagoon systems along the coastal plains of Rio Grande do Sul State. Sampling was performed in different freshwater bodies along the eastern (Casamento Lake area) and western (near the city of Tapes) banks of the Patos Lagoon (30º40' S-30º10' S and 50º30' W-51º30' W). The samples were collected once in the rainy season and once in the dry season (from May 2003 to December 2003) using a plankton net (25 µm mesh) in pelagic and littoral zones, and by squeezing the submerged parts of aquatic macrophytes. Thirty one species belonging to the families Synechoccocaceae (7 taxa), Merismopediaceae (12 taxa), Chamaesiphonaceae (1 taxon), Microcystaceae (4) and Chroococcaceae (7 taxa) were identified. Among these species, five are reported for the first time in Rio Grande do Sul State: Chamaesiphon amethystinus (Rostafinski) Lemmermann, Chroococcus minimus (Keissler) Lemmermann, Coelomoron pusillum (Van Goor) Komárek, Coelosphaerium kuetzingeanum Naegeli, and Cyanodictyon tubiforme Cronberg.

Cyanobacteria from coastal lagoons in southern Brazil: non-heterocytous filamentous organisms

This study describes and illustrates non-heterocytous filamentous cyanobacteria found in lagoon systems on the coastal plains of Rio Grande do Sul State. Collections were carried out in different freshwater bodies along the eastern (Casamento Lake area) and western (Tapes City area) margins of the Patos Lagoon (UTM 461948-6595095 and 542910-6645535) using a plankton net (25 µm mesh) in pelagic and littoral zones as well as by squeezing submerged parts of aquatic macrophytes, during both the rainy and dry seasons, from May to December/2003. Twenty two species belonging to the families Phormidiaceae (eight taxa), Pseudanabaenaceae (seven taxa), Oscillatoriaceae (six taxa), and Spirulinaceae (one taxon) were identified. Among these species, five are reported for the first time from Rio Grande do Sul State: Leptolyngbya cebennensis, Microcoleus subtorulosus, Oscillatoria cf. anguina, O. curviceps and Phormidium formosum.

Evidence for the presence of a kininogen-like species in a case of total deficiency of low and high molecular weight kininogens

Low and high molecular weight kininogens (LK and HK), containing 409 and 626 amino acids with masses of ~65 and 120 kDa after glycosylation, respectively, are coded by a single gene mapped to the human chromosome 3 by alternative splicing of the transcribed mRNA. The NH2-termini Glu1-Thr383 region, identical in LK and HK, contains bradykinin (BK) moieties Arg363-Arg371. LK, HK and their kinin products Lys-BK and BK are involved in several biologic processes. They are evolutionarily conserved and only 7 patients, all apparently normal, have been reported to lack them. In one of these patients (Williams' trait), a codon mutation (Arg178 ® stop) has been blamed for the absence of LK and HK. However, using Western blots with 2 monoclonal anti-HK antibodies, one that recognizes the region common to LK and HK and the other that recognizes only HK, I detected ~110-kDa bands in the plasma of this LK/HK-deficient patient vs ~120-kDa bands in normal human and ape plasmas. With polyclonal anti-Lys-BK antibody, which strongly detects BK cleaved at its COOH-terminus in purified HK, I detected ~110-kDa bands in the normal and the deficient plasmas. Western blots with a monoclonal anti-prekallikrein (PK) antibody showed that surface activation of PK and distribution of PK activation products, both dependent on HK, were similar in these plasmas. These findings suggest that a mutant gene yielded a kininogen-like species possibly involving aberrant mRNA splicing - structurally different from normal HK, but apparently with the capacity to carry out seemingly vital HK functions.

Anxiogenic-like effect of acute and chronic fluoxetine on rats tested on the elevated plus-maze

The selective serotonin reuptake inhibitor fluoxetine (FLX) is widely prescribed for depression and anxiety-related disorders. On the other hand, enhanced serotonergic transmission is known to be classically related to anxiety. In this study, the effects of acute (5.0 mg/kg) and chronic (5.0 mg/kg, 22 days) FLX were investigated in both food-deprived and non-deprived rats tested in the elevated plus-maze. Significant main effects of the three factors (drug, food condition and administration regimen) were observed, but no interaction between them. The administration of either acute or chronic FLX resulted in an anxiogenic effect, as detected by a significant reduction in the percentage of time spent in the open arms and in the percentage of open arm entries. Food deprivation yielded an anxiolytic-like profile, probably related to changes in locomotor activity. The administration regimen resulted in an anxiolytic profile in chronically treated rats, as would be expected after 22 days of regular handling. The anxiogenic action of acute FLX is consistent with both its neurochemical and clinical profile. The discrepancy between the anxiogenic profile of chronic FLX and its therapeutic uses is discussed in terms of possible differences between the type of anxiety that is measured in the plus-maze and the types of human anxiety that are alleviated by fluoxetine.

Antidepressant-like effect of lead in adult mice

It has been reported that lead can cause behavioral impairment by inhibiting the N-methyl-D-aspartate (NMDA) receptor complex. MK-801, a noncompetitive NMDA receptor antagonist, exhibits an antidepressant-like action in the forced swimming test. The purpose of the present study was to determine whether subacute lead exposure in adult male Swiss mice weighing 30-35 g causes an antidepressant-like action in a forced swimming test. Mice were injected intraperitoneally (ip) with 10 mg/kg lead acetate or saline daily for 7 consecutive days. Twenty-four hours after the last treatment, the saline and lead-treated mice received an injection of MK-801 (0.01 mg/kg, ip) or saline and were tested in forced swimming and in open-field tests. Immobility time was similarly reduced in the saline-MK-801, Pb-saline and Pb-MK-801 groups compared to the saline-saline group (mean ± SEM; 197.3 ± 18.5, 193.5 ± 15.8, 191.3 ± 12.3 and 264.0 ± 14.4 s, respectively; N = 9). These data indicate that lead may exert its effect on the forced swimming test by directly or indirectly inhibiting the NMDA receptor complex. Lead treatment caused no deficit in memory of habituation and did not affect locomotor activity in an open-field (N = 14). However, mice that received MK-801 after lead exhibited a deficit in habituation (22% reduction in rearing responses between session 3 and 1; N = 14) as compared to control (41% reduction in rearing responses; N = 15), further suggesting that lead may have affected the NMDA receptor activity. Forced-swim immobility in a basin in two daily consecutive sessions was also significantly decreased by lead exposure (mean ± SEM; day 1 = 10.6 ± 3.2, day 2 = 19.6 ± 3.6; N = 16) as compared to control (day 1 = 18.4 ± 3.8, day 2 = 34.0 ± 3.7; N = 17), whereas the number of crossings was not affected by lead treatment, further indicating a specific antidepressant-like action of lead.

Kallikrein-like amidase activity in renal ischemia and reperfusion

We assessed a kallikrein-like amidase activity probably related to the kallikrein-kinin system, as well as the participation of leukocyte infiltration in renal ischemia and reperfusion. Male C57BL/KSJmdb mice were subjected to 20 or 60 min of ischemia and to different periods of reperfusion. A control group consisted of sham-operated mice, under similar conditions, except for ischemia induction. Kallikrein-like amidase activity, Evans blue extravasation and myeloperoxidase activity were measured in kidney homogenates, previously perfused with 0.9% NaCl. Plasma creatinine concentration increased only in the 60-min ischemic group. After 20 min of ischemia and 1 or 24 h of reperfusion, no change in kallikrein-like amidase activity or Evans blue extravasation was observed. In the mice subjected to 20 min of ischemia, edema was evident at 1 h of reperfusion, but kidney water content returned to basal levels after 24 h of reperfusion. In the 60-min ischemic group, kallikrein-like amidase activity and Evans blue extravasation showed a similar significant increase along reperfusion time. Kallikrein-like amidase activity increased from 4 nmol PNA mg protein-1 min-1 in the basal condition to 15 nmol PNA mg protein-1 min-1 at 10 h of reperfusion. For dye extravasation the concentration measured was near 200 µg of Evans blue/g dry tissue in the basal condition and 1750 µg of Evans blue/g dry tissue at 10 h of reperfusion. No variation could be detected in the control group. A significant increase from 5 to 40 units of DAbs 655 nm g wet tissue-1 min-1 in the activity of the enzyme myeloperoxidase was observed in the 60-min ischemic group, when it was evaluated after 24 h of reperfusion. Histological analysis of the kidneys showed migration of polymorphonuclear leukocytes from the vascular bed to the interstitial tissue in the 60-min ischemic group after 24 h of reperfusion. We conclude that the duration of ischemia is critical for the development of damage during reperfusion and that the increase in renal cortex kallikrein-like amidase activity probably released from both the kidney and leukocytes may be responsible, at least in part, for the observed effects, probably through direct induction of increased vascular permeability.