Patrícia Schwarz

Development and Application of an Enzyme-Linked Immunosorbent Assay to Detect Antibodies against Prevalent Salmonella Serovars in Swine in Southern Brazil

The implementation of Salmonella control programs in the pork production chain demands rapid and cost-effective methods to assess the prevalence of infection in pig herds. The objective of the present study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) based on S. Typhimurium lipopolysaccharides (LPS) to measure the prevalence of infection caused by Salmonella in swine herds. Coating antigen was produced by phenol extraction of S. Typhimurium culture. After standardization of ELISA test conditions, the assay was validated by testing serum samples on different animal categories: pigs orally inoculated with S. Typhimurium and sentinel animals in contact with them, naturally infected animals, colostrum-deprived piglets, and bacterin-immunized pigs. Seroconversion was observed in inoculated pigs (7 days postinfection [DPI]) and in the sentinels (21 DPI). Nonspecific reactions were not detected in the sera of colostrum-deprived animals. Serum samples from animals immunized with Salmonella Agona, Salmonella Derby, Salmonella Panama, and Salmonella Bredeney bacterins showed marked cross-reaction with the LPS from the serovar Typhimurium. Moreover, positive results obtained with the in-house ELISA were associated with Salmonella isolation in 75 infected pig herds. Comparisons with 2 commercial kits showed a linear correlation coefficient of 0.847 between the in-house ELISA and kit A and 0.922 with kit B but a low agreement in the qualitative results. In conclusion, the newly developed in-house ELISA based on S. Typhimurium LPS can be a useful tool to determine the intensity of Salmonella sp. infection in swine herds.

Presença de Salmonella sp. no trato intestinal e em tonsilas/linfonodos submandibulares de suínos ao abate

The association between Salmonella sp. isolated from the intestinal tract of pigs at slaughter and its presence in tonsils/mandibular lymph nodes (T/MDL) used in the sausage manufacture was assessed. In a trial mesenteric lymph nodes (LM), intestinal contents (IC) and T/MDL were collected from 90 animals. Salmonella sp. was isolated from 71 (79%) animals in at least one of the samples. T/MDL contaminated samples were correlated with Salmonella sp. isolated from LM and/or IC (P<0.001), with odds ratioLTpositive 5.6. In other trial, T/MDL of 75 animals processed separately, were associated with isolation of Salmonella sp (P< 0.0001). The serovars more prevalent were Typhimurium and Bredeney. It was concluded that Salmonella sp. presence in the intestinal tract is a risk factor for contamination of the portions of carcasses used in sausage manufacture.

Salmonella enterica: isolamento e soroprevalência em suínos abatidos no Rio Grande do Sul

The prevalence of Salmonella in slaughtered swines was evaluated and the results of serology and Salmonella isolation were compared to determine the most important stage of the infection (on-farm or transport and pre-slaughter) in relation to the number of carrier animals found at slaughter. Forty herds from three different swine raisers of the southern region of Brazil were sampled at slaughter. Blood and mesenteric lymph nodes were collected from 20 swines of each herd. Serum samples were submitted to ELISA including somatic antigens of S. Typhimurium. Salmonella was isolated from 62.5% to 85.0% of the animals, while seroprevalence varied from 73.8% to 83.2% in the three swine raisers. Serovar Agona, Typhimurium, and Panama were the most prevalent among the Salmonella isolates. Isolation and serological results demonstrated that Salmonella infection occurred during the on-farm stage, since a high seroprevalence was detected at slaughter. It was concluded that a Salmonella control program in southern Brazil must be started with the implementation of intervention measures at farm level.

Extracorporeal membrane oxygenation for postpneumonectomy ARDS

A SARA pos-pneumonectomia e uma complicacao infrequente, porem com alta mortalidade (de 50% ate 100%).(1) Gostariamos de relatar um caso manejado satisfatoriamente com extracorporeal membrane oxygenation (ECMO, oxigenacao extracorporea por membrana). Uma mulher branca de 31 anos, com diagnostico de fibrose cistica havia 10 anos, apresentava pneumonias repetidas e secundarias a bronquiectasias que predominavam no pulmao esquerdo (Figura 1A). Nos ultimos dois anos, apesar do uso ininterrupto de antibiotico

Association between vitamin D levels and inflammatory activity in brain death: A prospective study

BACKGROUND Vitamin D insufficiency is linked to several common inflammatory disorders. Brain death (BD) causes a massive catecholamine release, leading to intense inflammatory activity. We aimed to evaluate vitamin D serum levels in brain-dead individuals in comparison to critically ill patients without BD to assess the correlation between vitamin D and cytokine levels. METHODS Sixteen brain-dead patients and 32 critically ill controls were prospectively enrolled. Blood samples from 25 brain-dead patients from a previous study were also used for vitamin D quantification. Plasma TNF, IL-1β, IL-6, IL-8, IL-10, IFN-γ and serum vitamin D levels were compared using Students t-test or one-way ANOVA. Spearmans test was used to assess the correlation between vitamin D and cytokine levels. RESULTS Mean vitamin D levels were 16.4 ± 7.9 ng/mL, with 52 patients (71.2%) classified as vitamin D deficient (serum levels < 20 ng/mL). Vitamin D levels were similar in 41 brain-dead patients as compared to control subjects (15.6 ± 6.9 ng/mL vs 17.4 ± 9.0 ng/mL; p = 0.383). Moderate direct correlations were observed between vitamin D and IL-8, IL-10, and IFN-γ in the prospective group of 16 brain-dead patients (IL-8: r = 0.5, p = 0.049; IL-10 r = 0.67, p = 0.005; IFN-γ r = 0.6, p = 0.015). Vitamin D was inversely correlated with IL-6 (r = -0.36, p = 0.044) in critically ill controls. CONCLUSIONS Vitamin D serum levels were similarly low in brain-dead and critically ill patients. In brain-dead patients, vitamin D serum levels correlated with plasma IL-8, IL-10 and IFN-γ.

Brain Death-Induced Inflammatory Activity is Similar to Sepsis-Induced Cytokine Release

Brain death (BD) is associated with a systemic inflammation leading to worse graft outcomes. This study aimed to compare plasma cytokine values between brain-dead and critically ill patients, including septic and non-septic controls, and evaluate cytokine release kinetics in BD. Sixteen brain-dead and 32 control patients (16 with and 16 without sepsis) were included. Plasma cytokines were measured by magnetic bead assay after the first clinical exam consistent with BD and every 6 hours thereafter, and at the time of study entry in the control group. The values for IL-8 and IFN-γ were higher in brain-dead and septic patients than in non-septic patients [IL-8: 80.3 (18.7–169.6) vs. 68.2 (22.4–359.4) vs. 16.4 (9.2–42.7) pg/mL; P = 0.006; IFN-γ: 2.8 (1.6-6.1) vs. 3.4 (1.2–9.0) vs. 0.5 (0.5–1.8) pg/mL; P = 0.012]. TNF showed a clear tendency to increase in brain-dead patients [2.7 (1.0–4.8) vs. 1.0 (1.0–5.6) vs. 1.0 (1.0–1.0) pg/mL; P = 0.051], and IL-6 values were higher in brain-dead patients than in non-septic controls [174.5 (104.9–692.5) vs. 13.2 (7.3–38.6) pg/mL; P = 0.002]. These differences remained even after excluding brain-dead patients who also had sepsis (n = 3). IL-1β and IL-10 values increased from baseline to time point 2 (∼6 hours later) [IL-1β: 5.39 (1.93–16.89) vs. 7.11 (1.93–29.13) pg/mL; P = 0.012; IL-10: 8.78 (3.62–16.49) vs. 15.73 (5.49–23.98) pg/mL; P = 0.009]. BD-induced and sepsis-induced plasma cytokine values were similarly high, and both were higher than the observed in non-septic critically ill patients.

Viabilidade espermática após a troca de plasma seminal entre . machos suínos com diferente sensibilidade ao resfriamento

O objetivo deste estudo foi verificar um possivel efeito do plasma seminal (PS) na sensibilidade espermatica ao resfriamento,a partir da troca cruzada de PS, entre cachacas com diferente mahutencao da motilidade. (MOT) a 17°C. Foram utilizados comodoadores de semen cinco cachacas selecionados pre.viamente, de um total de 30 machos, conforme a MOT do semenavaliada durante sete dias de armazenamento a 17°C, e classificados como: menor (MES - MOT 360% apos 144h) e maiorsensibilidade ao resfriamento (MAS- MOT <60% nas 72h). Ejaculados dos machos MAS e MES foram distribuidos em seistratamentos, com cinco repeticoes cada. Nos tratamentos T1 e T3, o semen dos. machos MES e MAS, respectivamente, foramprocessados de acordo com o protocolo convencional. Foi efetuada centrifugacao (800g por 1 O min) e adicao do PS (1 Oml)homologo para os espermatozoides MES eMAS, respectivamente, nos T2 e T4. Apos a centrifugacao, foi realizada a troca doPS, sendo que espermatozoides dos machos MES foram expostos ao ps dos machos MAS (T5) e o PS dos machos MES foiadicionado aos espermatozoides dos machos MAS (T6). As doses de semen; diluidas em BTS, contendo tres bilhoes deespermatozoides e distribuidas· de acordo com os tratamentos descritos foram avaliadas durante sete dias de conservacao.A MOT foi avaliada a cada .24h. Os percentuais de acrossomas normais (NAR) e de membranas integras (IM) foram avaliadosnas 24, 72, 120 e 168h. Diferencas significativas (P= 0,038) na MOT, entre os machos MES (T1) e MAS (T3), foram observadasapos armazenamento de 48 horas. Em termos de IM, foi verificado um maior percentual (P= 0,027), para os machos MES, naavaliacao efetuada nas 12O horas. Diferencas significativas (P<0;01) no NAR; foram ·tambem verificadas, para os machos MAS,embora somente nas 72 horas, quando o ejaculado desses machos foi· submetido a centrifugacao e adicao de PS homologoou heterologo. Nao foram observadas alteracoes na MOT e na IM, quando foi efetuada a troca de PS entre os machos MES eMAS. Os espermatozoides oriundos dos machos MAS apresentaram diminuicao de 11% na MOT nas 24 horas, quandosubmetidos a centrifugacao, ao passo que espermatozoides oriundos dos machos MES apresentaram diminuicao de 16% naMOT (P= 0,054), porem somente nas 168 horas. Uma diminuicao da IM, embora somente nas 168 horas, foi tambem observada,tanto para os machos MES como para os MAS (P<0,01 ), quando o semen destes machos foi centrifugado e adicionadode PS homologo ou heterologo. A centrifugacao, utilizada durante o processamento de troca do PS, entre machos comdiferente sensibilidade ao resfriamento, tem efeitos deleterios sobre a motilidade e a integridade de membrana da celulaespermatica, sobretudo para os machos MAS. Nao foi possivel reverter a maior sensibilidade ao resfriamento de espermatozoidessuinos, apos a ejaculacao, com a adicao de .1 O% do PS de machos com semen de menor sensibilidade.