Clonagem, expressão e diversidade na seqüência do gene iss de Escherichia coli patogênica para aves (APEC), isolada no Brasil

The Iss (Increased serum survival) protein is an important characteristic of resistance to complement system of avian pathogenic Escherichia coli (APEC). The objectives of this work were to cloning and verify the sequence diversity of iss gene from APEC and characterize the recombinant Iss protein. The iss gene of 309 bp was amplified by PCR, cloned and expressed in E. coli BL21 (DE3) using the pET SUMO vector. The iss gene from APEC9 strain was classified as iss type 1 by differentiation of the three iss gene allele types. The protein was expressed by induction of IPTG and purified in resin charged with the nickel ion. Antibodies IgY anti rIss reacted with rIss showing a molecular mass of 22 kDa, corresponding 11KDa of Iss protein and 11 KDa SUMO protein.A proteína Iss (increased serum survival) é uma importante característica de resistência ao sistema complemento da Escherichia coli patogênica para aves (APEC). Os objetivos deste trabalho foram clonar e verificar a diversidade da seqüência do gene iss de APEC e caracterizar a proteína Iss recombinante. O gene iss de 309 bp foi amplificado por PCR, clonado e expresso na E. coli BL21 (DE3) utilizando o vetor pET SUMO. O gene iss da APEC9 foi classificado como iss tipo 1 pela diferenciação entre 3 tipos de iss alelos. A proteína Iss foi expressa pela indução com IPTG, purificada em coluna com resina ligada ao íon níquel e utilizada na imunização de galinhas poedeiras. Anticorpos da classe IgY anti rIss reagiram com a proteina rIss, a qual apresentou massa molecular de 22 kDa, correspondendo 11kDa da Iss e 11 kDa da proteína SUMO

Evaluation of potential immunostimulant of the Carboxymethyl-glucan from Saccharomyces cerevisiae in poultry (Gallus domesticus)

A carboximetilglucana (CMG) é uma molécula solúvel, composta de resíduos de glicopiranosil unidos em ?(1-3) e ?(1-6), que possui a capacidade de ativar o sistema imune do hospedeiro. O objetivo do presente estudo foi avaliar as características produtivas e imunológicas de 192 frangos de corte (Gallus domesticus) da linhagem COBB, que receberam rações contendo 0%, 0,025%, 0,050% e 0,075% de CMG de Saccharomyces cerevisiae adicionada em farinha de milho.  Todas as aves foram imunizadas contra a doença de Newcastle e, em cada tratamento, 3 aves escolhidas aleatoriamente receberam CMG intramuscular no 3º, 7º e 14º dia. Foram avaliados o desempenho animal, o desenvolvimento da bursa de Fabricius e lâminas histológicas do intestino delgado, além do número de células fagocíticas no sangue e níveis de anticorpos no soro. Os resultados mostraram diferença no ganho de peso e consumo de ração das aves que consumiram a CMG no período de 1 a 21 dias. O peso relativo da bursa de Fabricius aumentou nas aves suplementadas com 0,025 e 0,050% de CMG. O número de células fagocíticas e os níveis de anticorpos totais dos frangos de corte, aos 21 dias de idade foram maiores nas aves que receberam CMG na ração. Para os animais que receberam CMG via intramuscular foi observado aumento no título dos anticorpos específicos para Newcastle.The carboxymethylglucan (CMG) is a soluble molecule, composed of glucopyranosyl linked by ?(1-3) e ?(1-6), which can activate the immune system of the host. The purpose of this study was evaluate the productive and immunological characteristics of 192 poultry (Gallus domesticus) COBB line which received feds containing 0%, 0,025%, 0,050% e 0,075% of CMG from Saccharomyces cerevisiae added in corn flour. All poultry were immunized against Newcastle disease and at each treatment 3 poultries randomly chosen received CMG intramuscular at 3, 7 and 14 days. It was evaluated the animal performance, development of the bursa of Fabricius, histological slides of the small intestine, counts of  phagocytes cells in blood and levels of antibodies in serum. The results showed difference in weight gain and consumption of feed to poultry that consumed CMG at 1 to 21 days. Fabricius bursa relative weight increased in poultry supplemented with 0,025 e 0,050% of CMG. The phagocytic cells number and total levels of antibodies found in poultry at 21 days were higher in those that received CMG in the diet. For the animals that received intramuscular CMG was observed increase of antibodies specific to Newcastle

Anti-dentine Salivary SIgA in young adults with a history of dental trauma in deciduous teeth

Anti-dentin autoantibodies are associated with inflammatory root resorption in permanent teeth and are modulated by dental trauma and orthodontic force. However, it is not known whether deciduous tooth trauma can stimulate the development of a humoral immune response against dentin. The aim of this study was to evaluate the levels of salivary SIgA reactivity against human dentin extract in young adults with a history of trauma in the primary dentition. A sample of 78 patients, aged 18 to 25, who had completed an early childhood (0 to 5 years old) caries prevention program years earlier at the Universidade Estadual de LondrinaPediatric Clinic, underwent radiographic examination and salivary sampling. Anti-dentin SIgA levels were analyzed by immunoenzymatic assay and Western blotting. Although dental trauma to deciduous teeth had occurred in 34 (43.6%) of the patients, no differences in SIgA levels were detected between individuals who had experienced trauma and those who had not (p > 0.05). Multivariate regression analysis showed no association between dental trauma and SIgA levels (p > 0.05). Patients with a history of deciduous trauma presented low levels of anti-dentin antibodies, associated with orthodontic root resorption (p < 0.05). Western blot analysis showed that salivary antibodies recognized a single band of approximately 45 kDa in dentin extract. We concluded that salivary SIgA recognizes a specific component of the dentin matrix and that anti-dentin antibodies were not triggered by trauma to primary teeth. However, trauma to deciduous teeth may down-modulate SIgA in response to orthodontic root response.</p

Metallo-β-lactamase-production in meropenem-susceptible Pseudomonas aeruginosa isolates: risk for silent spread

The aim of this study was to characterize two metallo-β-lactamases (MBLs)-producing Pseudomonas aeruginosa clinical isolates showing meropenem susceptibility. Antimicrobial susceptibility was assessed by automated testing and Clinical and Laboratory Standards Institute agar dilution method. MBL production was investigated by phenotypic tests. Molecular typing was determined by pulsed field gel electrophoresis (PFGE). MBL-encoding genes, as well as their genetic context, were identified by polymerase chain reaction (PCR) and sequencing. The location of blaIMP-16 was determined by plasmid electrophoresis, Southern blot and hybridization. Transcriptional levels of blaIMP-16, mexB, mexD, mexF, mexY, ampC and oprD were determined by semi-quantitative real time PCR. The P. aeruginosa isolates studied, Pa30 and Pa43, showed imipenem and meropenem susceptibility by automated testing. Agar dilution assays confirmed meropenem susceptibility whereas both isolates showed low level of imipenem resistance. Pa30 and Pa43 were phenotypically detected as MBL producers. PFGE revealed their clonal relatedness. blaIMP-16 was identified in both isolates, carried as a single cassette in a class 1 integron that was embedded in a plasmid of about 60-Kb. Pa30 and Pa43 overexpressed MexAB-OprM, MexCD-OprJ and MexXY-OprM efflux systems and showed basal transcriptional levels of ampC and oprD. MBL-producing P. aeruginosa that are not resistant to meropenem may represent a risk for therapeutic failure and act as silent reservoirs of MBL-encoding genes

Are DMI+QoI premixes applied during flowering worth for protecting wheat yields from Fusarium head blight? A meta-analysis

Fusarium head blight (FHB), caused mainly by Fusarium graminearum, is best controlled with demethylation inhibitor (DMI) fungicides applied during flowering. However, the use of premixes of DMI and quinone outside inhibitor (QoI) fungicides to control FHB has increased in Brazil, but the individual results are inconsistent. Data on FHB severity and wheat yields measured in field experiments conducted in Brazil were gathered from both peer- and non-peer-reviewed sources published from 2000 to 2018. After applying selection criteria, 35 bibliographic sources, contributing 73 (50% from cooperative trials) trials, were identified. At least one of four DMI+QoI premixes and one tebuconazole (TEB) treatment, applied mostly twice (full-flowering and 10 days) tested in at least 14 trials and three year each, were present in a selected trial. Estimates of percent control (and respective 95%CI) by a network model ranged from 44.1% (pyraclostrobin + metconazole applied once; 32.4 - 53.7) to 64.3% (pyraclostrobin + metconazole; 58.4 - 69.3); the latter not differing from TEB (59.9%, 53.6 - 65.3). Yield response was statistically similar for pyraclostrobin + metconazole (532.1 kg/ha, 441 - 623) and trifloxystrobin + prothioconazole (494.9 kg/ha, 385 - 551), and both differed statistically from a group composed of TEB (448.2 kg/ha, 342 - 554), trifloxystrobin + TEB (468.2 kg/ha, 385 - 551), azoxystrobin + TEB (462.4 kg/ha, 366 - 558) and pyraclostrobin + metconazole applied once (413.7 kg/ha, 308 - 518). The two categories of FHB index (7% cut off) and yield (3,000 kg/ha), both in the non-treated check, did not explain the heterogeneity in the estimates. The probability of not-offsetting control costs was generally lower than 0.45 for scenarios considering two sequential sprays of the low-cost TEB or one spray of pyraclostrobin + metconazole as management choices. The envisioned enhanced economic return, solely based on yield response, from using two sprays of DMI+QoI premixes to control FHB should be seen with caution given the marginal levels of profitability

Spray-dried plasma in the recovery of light piglets at weaning: production performance, blood profile, diarrhea frequency and economic viability

O objetivo desse trabalho foi avaliar o desempenho, perfil hematológico, freqüência de diarréia e potencial econômico de uso do plasma sanguíneo desidratado (PSD) na dieta de leitões desmamados com baixo peso, aos 21 dias de idade. Foram utilizados 72 leitões, sendo 48 machos castrados e 24 fêmeas (PenArLan) com idade média inicial de 21 dias. O delineamento experimental foi de blocos ao acaso, com 4 tratamentos e 6 repetições por tratamento (cada baia com dois machos e uma fêmea foi considerada uma repetição). Os tratamentos foram: T1 - animais pesados ao desmame, sem suplementação; T2 - animais leves ao desmame, suplementados com 10g/animal/dia de PSD; T3 - animais leves ao desmame, suplementados com 20g/animal/dia de PSD; T4 - animais leves ao desmame, sem suplementação. O PSD (10 e 20g/animal/dia) foi fornecido diretamente no cocho três vezes ao dia. Aos 35 e 70 dias de idade os leitões foram submetidos a coleta de sangue, sendo utilizado 8 animais de cada tratamento. Diariamente foi realizada a avaliação da freqüência de diarréia. Foi verificada diferença para conversão alimentar (P 0,05) para freqüência de diarréia. Houve um melhor resultado na análise econômica, apontando vantagem para o T3 em relação ao custo da ração nos períodos compreendidos entre 21 a 35 dias, 36 a 49 dias e entre 21 a 49 dias de idade. Considerando todo o período experimental (21 a 70 dias de idade) o T3 apresentou melhor índice de eficiência econômica e de custo médio.The aim of this research was to evaluate the effects of spray-dried plasma (SDP) on the performance, hematological profile, diarrhea frequency and economic viability of this ingredient in diets for light piglets at weaning, with 21 days of age. The trial was done using 72 pigs, 48 castrated male and 24 female (PenArLan), with an initial age of 21 days. The experimental design consisted of randomized blocks, with 4 treatments and 6 repetitions (each pen with two males and one female was considered a repetition). The treatments were: T1 - heavy weight weaned pigs, without supplementation; T2 - light weight weaned pigs, supplemented with 10g/animal/day of SDP; T3 - light weight weaned pigs, supplemented with 20g/animal/day of SDP; T4- light weight weaned pigs, without supplementation. The SDP were fed daily directly at the feeder three times per day. At 35 and 70 days old piglets were subjected to blood collection, and 8 animals in each treatment were used. Daily evaluation was made of diarrhea frequency. There was difference to feed:gain (P 0,05) on frequency of diarrhea. There were differences in feed cost with advantage to T3 in the period of 21 to 35; 36 to 49 and 21 to 49 days of age. In total period (21 to 70 days of age) T3 showed the best index of economic efficiency