Existence of SARS-Cov-2 in the Peritoneal Fluid / Existência de SARS-Cov-2 no líquido peritoneal

Abstract Objective To determine the existence of SARS-CoV-2 in the peritoneal fluid to assess the risk of exposure through surgical smoke and aerosolization threatening healthcare workers during abdominal surgery. Background SARS-CoV-2 is a respiratory virus and possible ways of viral transmission are respiratory droplets, close contact, and fecal-oral route. Surgeries pose risk for healthcare workers due to the close contact with patients. Aerosolized particles may be inhaled via the leaked CO2 during laparoscopic procedures and surgical smoke produced by electrocautery. Methods All the data of 8 patients, who were tested positive for COVID-19, were collected between August 31, 2020 and April 30, 2021. Recorded clinicopathologic data included age, symptoms, radiological and laboratory findings, antiviral treatment before surgery, type of surgery and existence of the virus in the peritoneal fluid. Nasopharyngeal swab RT-PCR was used for the diagnosis. COVID-19 existence in the peritoneal fluid was determined by RT-PCR test as well. Results All 8 COVID-19 positive patients were pregnant, and surgeries were cesarean sections. 1 of the 8 patients was febrile during surgery. Also only 1 patient had pulmonary radiological findings specifically indicating COVID-19 infection. Laboratory findings were as follows: 4 of 8 had lymphopenia and all had elevated D-dimer levels. Peritoneal and amniotic fluid samples of all patients were negative for SARS-CoV-2. Conclusion SARS-CoV-2 exposure due to aerosolization or surgical fumes does not seem to be likely, provided the necessary precautions are taken.

Resumo Objetivo Determinar a existência de SARS-CoV-2 no fluido peritoneal para avaliar o risco de exposição através da fumaça cirúrgica e aerossolização que ameaçam os profissionais de saúde durante a cirurgia abdominal. Contexto O SARS-CoV-2 é um vírus respiratório e as possíveis formas de transmissão viral são gotículas respiratórias, contato próximo e rota fecal-oral. As cirurgias representam risco para os profissionais de saúde devido ao contato próximo com os pacientes. As partículas aerossolizadas podem ser inaladas através do CO2 vazado durante os procedimentos laparoscópicos e a fumaça cirúrgica produzida pela eletrocauterização. Métodos Todos os dados de 8 pacientes, que foram testados positivos para COVID-19, foram coletados entre 31 de agosto de 2020 e 30 de abril de 2021. Dados clinicopatológicos registrados incluíam idade, sintomas, achados radiológicos e laboratoriais, tratamento antiviral antes da cirurgia, tipo de cirurgia e existência do vírus no fluido peritoneal. O diagnóstico foi feito através do swab nasofaríngeo RT-PCR. A existência de COVID-19 no fluido peritoneal foi determinada pelo teste de RT-PCR também. Resultados Todas as 8 pacientes positivas para COVID-19 estavam grávidas, e as cirurgias eram cesarianas. 1 das 8 pacientes estava com febre durante a cirurgia. Também apenas 1 paciente tinha achados radiológicos pulmonares especificamente indicando infecção por COVID-19. Os achados laboratoriais foram os seguintes: 4 de 8 tinham linfopenia e todas apresentavam níveis elevados de D-dímero. Amostras de fluido peritoneal e líquido amniótico de todas as pacientes foram negativas para SARS-CoV-2. Conclusão A exposição ao SARS-CoV-2 devido à aerossolização ou fumaças cirúrgicas não parece ser provável, desde que sejam tomadas as precauções necessárias.

The anatomical site of perforation peritonitis and their microbiological profile: a cross-sectional study

Background: Initiation of early appropriate antibiotic therapy influences the outcome of perforation peritonitis, which otherwise is delayed till culture reports are available. The knowledge of microbial profile and sensitivity of peritoneal fluid culture with respect to the anatomical site of perforation peritonitis will help in initiation of early appropriate antibiotic therapy in the post-operative period.Methods: A cross-sectional study conducted from January 2017 to December 2017 where intraoperative peritoneal fluid sample in patients of perforation peritonitis was subjected to culture (aerobic and anaerobic) and sensitivity and results analysed with respect to anatomical site of perforation.Results: 50 patients were studied. The most common site of perforation was ileum (32%) followed by appendix (18%) and stomach (18%). In aerobic culture, the culture positivity rate was highest in colonic perforation (100%) and least in gastric perforation (44.4%). The most common organism isolated in all sites of perforation peritonitis was E. coli followed by Klebsiella spp. In anaerobic culture, although facultative anaerobes were isolated, no strict anaerobe was isolated. The most sensitive antibiotics covering all isolated organisms were gentamycin (p=0.006), colistin (p=0.018), piperacillin and tazobactum (p=0.022).Conclusions: The predominant differential normal flora according to site of gastrointestinal tract was not reflected in the peritoneal fluid culture of patients with perforation peritonitis and E. coli was the most common organism isolated in all sites of perforation peritonitis. The antibiotic sensitivity profile showed the increasing resistance against third generation cephalosporins. Aminoglycosides, piperacillin and tazobactum, meropenem and colistin showed a significant antimicrobial activity against organisms isolated from cases of perforation peritonitis.

Peritoneal fluid collection in healthy cattle and evaluation of changes in cell morphology during storage in refrigerated and non-refrigerated samples / Colheita do fluido peritoneal de bovinos sadios e verificação das alterações na morfologia celular quando mantido refrigerado ou à temperatura ambiente

This study aimed to evaluate the appropriate sites of abdominocentesis for peritoneal fluid collection in cattle and to investigate the time of cell viability in vitro, comparing three methods of sample conservation. Twenty-one healthy cattle (19 females and 2 males) were subjected to a laparocentesis procedure to obtain peritoneal fluid, with punctures in three defined sites: left cranial, right cranial, and right caudal. The total peritoneal fluid collected was divided into three aliquots and maintained under three preservation conditions: room temperature (26°C), refrigeration (4°C), and room temperature (26°C) with the addition of 1µL of 10% formaldehyde per 1mL of peritoneal fluid. The peritoneal fluid analysis performed immediately after collection consisted of: physical examination (color, appearance, volume, and specific gravity), biochemical measures (pH, total protein, fibrinogen, creatinine, and glucose), and cellularity (total and differential counts). The determination of proteins and the examination of cells were repeated in each separate aliquot at two, four, six, and eight hours after harvest. Data were analyzed through repeated measures ANOVA or Friedman test. The harvest was productive in 67% of cattle. The left cranial and the right cranial puncture sites were the most appropriate. Peritoneal fluid analyzed after collection, the total protein concentration ranged from 1.4 to 3.6g/dL, and number of leukocytes ranged from 54 to 1,322 cells/µL; 60 to 95% of leukocytes were lymphocytes. The protein concentration decreased, but the absolute values of leukocytes, lymphocytes, and segmented neutrophils did not change up to eight hours after collection, independent of the maintenance method. Cell lysis was delayed by cooling, and the addition of formaldehyde did not help preserve the integrity of cellular morphology. Laparocentesis is a safe and secure procedure in cattle and maybe more productive when performed in specific sites on the left or right sides of the cranial abdominal wall. Peritoneal fluid samples may be analyzed with reliable results for up to eight hours after collection when kept refrigerated and for up to six hours when kept at room temperature.(AU)

O estudo teve como objetivo avaliar os locais adequados de laparocentese para a colheita de fluido peritoneal de bovinos e estabelecer o tempo de viabilidade celular in vitro, comparando três métodos de conservação. Vinte e um bovinos hígidos (19 fêmeas e 2 machos) foram submetidos ao procedimento de laparocentese para obtenção de fluido peritoneal, com punção em três pontos definidos: cranial esquerdo, cranial direito e caudal direito. O volume total do líquido peritoneal foi dividido em três alíquotas mantidas sob três métodos de conservação: temperatura ambiente (26°C); refrigeração (4°C); e temperatura ambiente (26°C) com adição de 1µL de formol 10% para cada 1mL de líquido peritonial. A análise do líquido peritoneal realizada imediatamente após sua obtenção consistiu em: exames físico (cor, aspecto, volume e densidade); bioquímicos (pH, proteína total, fibrinogênio, creatinina e glicose); e da celularidade (contagens total e diferencial). A determinação de proteínas e o exame da celularidade foram repetidos, em cada alíquota separada, as duas, quatro, seis e oito horas após a colheita. Análise de variâncias de medidas repetidas ou teste de Friedman foram empregados para avaliação ao longo do tempo. A colheita foi produtiva em 67% dos bovinos e os locais de punção craniais esquerdo e direito foram os mais adequados. A concentração de proteína total variou de 1,4 a 3,6g/dL e o número de leucócitos de 54 a 1.322 células/µL, com predomínio de linfócitos (60 a 95% das células) no fluido peritoneal analisado logo após a colheita. A concentração de proteínas diminuiu, mas os valores absolutos de leucócitos, de linfócitos e de neutrófilos segmentados não se modificaram até oito horas após a colheita, independente do método de manutenção das amostras. A lise celular foi retardada pela refrigeração e a adição de formol não contribuiu para preservar a integridade da morfologia celular. A laparocentese é um procedimento seguro e de execução fácil em bovinos sendo mais produtiva quando realizada em locais específicos à esquerda ou à direita craniais da parede abdominal. Amostras de fluido peritoneal podem ser analisadas com resultados confiáveis quando mantidas refrigeradas por até oito horas após a colheita e quando mantidas à temperatura ambiente por até seis horas.(AU)

Peritoneal Fluid Analysis Clinicocytological Study

Introduction: Appropriate peritoneal fluid analysis is themost efficient and effective method of diagnosing the causeof peritoneal effusion. Both nonmalignant and malignantcauses of effusion can be identified by cytology in severalcases in correlation with clinical history and examination.With this basis, the present study on cytology of peritonealeffusion was taken up. Current research aimed to study thecytology of the peritoneal fluid in various diseases to establishclinicocytological correlation, for proper management ofpatient.Material and Methods: 115 samples of peritoneal fluidwere subjected to physical, biochemical and cytologicalexamination.Results: Peritoneal effusion was seen in 62.61% of femalesand 37.39% of males. 66.96% samples were transudative and33.04% were exudative. TLC was less than 500 cells/cu.mm inmost (74.02%) of transudative effusions. 47.36% of exudativeeffusion had TLC greater than 1000 cells/cu.mm and 39.47%of exudative effusion had TLC between 500-1000 cells/cumm. 95 (82.60%) samples had predominantly lymphocytes.18.26% of peritoneal effusions were positive for malignantcells. Most (85.71%) of malignant effusions were exudative.Primary site could be assessed by cytological examination in(57.14%) of malignant effusions.Conclusion: Cytological study of body effusions is neither ascreening test nor a method of early diagnosis of cancer. It isin fact a complete diagnostic modality which aims at pointingout the etiology of effusion as well as in certain cases a meansof prognostication of the disease process. Non malignantcauses are the more common causes of peritoneal effusion.Metastatic carcinomas are the most common tumors found ineffusions.

Babesia spp. no líquido peritoneal em cão com ascite - relato de caso / Babesia spp. in the peritoneal fluid in dog with ascites - case report

Babesia canis é um protozoário cosmopolita que parasita eritrócitos de cães domésticos e selvagens. O diagnóstico é realizado mediante a observação direta do microrganismo em hemácias no esfregaço de sangue periférico, métodos sorológicos e técnicas moleculares. O objetivo deste trabalho é relatar pela primeira vez a presença de merozoítos de Babesia spp. no líquido peritoneal de um cão com ascite. No Hospital Veterinário da Universidade Federal de Viçosa, foi atendido um cão, macho, sem raça definida, de sete meses de idade, com histórico de emaciação, apatia e abaulamento abdominal. No exame físico, foram evidenciadas mucosas hipocoradas, ascite, sopro sistólico grau IV/V e taquipneia. Nos exames laboratoriais, evidenciou-se anemia normocítica/normocrômica, trombocitopenia e hipoproteinemia. No esfregaço sanguíneo, foram observadas estruturas intraeritrocitárias compatíveis com Babesia spp. A avaliação do líquido ascítico foi compatível com transudato modificado e observaram-se inúmeras estruturas intra e extracelulares compatíveis com merozoítas de Babesia spp. A presença de microrganismos intra e extracelular poderia estar relacionada a uma lesão no baço com extravasamento do conteúdo para a cavidade abdominal. A coleta do líquido peritoneal pode ser uma alternativa para o diagnóstico de babesiose quando o animal com suspeita da infecção apresentar ascite.(AU)

Babesia canis is a cosmopolitan protozoan that parasites erythrocytes of domestic and wild dogs. The diagnosis is performed by direct observation of the microorganism in red blood cells in the peripheral blood smear, serological methods and molecular techniques. The aim of this work is to report for the first time the presence of merozoites of Babesia spp. in the peritoneal fluid of a dog with ascites. At the Veterinary Hospital of the Federal University of Viçosa was attended a Mixed-breed seven month old dog, male, with history of emaciation, apathy and abdominal bulging. Pale mucous membranes, ascites, grade IV/V systolic murmur and tachypnea were evidenced in the physical examination. Laboratory tests revealed normocytic/normochromic anemia, thrombocytopenia, and hypoproteinemia. Intra-erythrocyte structures compatible with Babesia spp. were observed in the blood smear. The evaluation of the ascites fluid was compatible with modified transudate where numerous intra and extracellular structures compatible with Babesia spp. merozoites were observed. The presence of intra and extracellular microorganisms could be related to an injury of the spleen with extravasation of the contents into the abdominal cavity. Collection of the peritoneal fluid may be an alternative for the diagnosis of babesiosis when the animal with suspected infection has ascites.(AU)

Clinical significance of carcinoembryonic antigen in peritoneal fluid detected during operation in stage I–III colorectal cancer patients

BACKGROUND/AIMS: Early diagnosis of peritoneal metastases in patients with colorectal cancer (CRC) can influence patient prognosis. The aim of this study was to identify the clinical significance of carcinoembryonic antigen (CEA) in peritoneal fluid detected during operation in stage I–III CRC patients. METHODS: Between April 2009 and April 2015, we reviewed medical records from a total of 60 stage I–III CRC patients who had peritoneal fluid collected during operation. Patients who had positive cytology in the assessment of peritoneal fluid were excluded. We evaluated the values of CEA in peritoneal fluid (pCEA) to predict the long-term outcomes of these patients using Kaplan-Meier curves and Cox regression models. RESULTS: The median follow-up duration was 37 months (interquartile range, 21–50 months). On receiver operating characteristic analysis, pCEA had the largest area under the curve (0.793; 95% confidence interval, 0.635–0.950; P=0.001) with an optimal cutoff value of 26.84 (sensitivity, 80.0%; specificity, 76.6%) for predicting recurrence. The recurrence rate was 8.1% in patients with low pCEA ( < 26.84 ng/mL, n=37), and 52.2% in patients with high pCEA (≥26.84 ng/mL, n=23). In multivariate Cox regression analysis, high pCEA (≥26.84 ng/mL) was a risk factor for poor cancer-free survival (CFS) in stage I–III patients. CONCLUSIONS: In this study, we determined that high pCEA (≥26.84 ng/mL) detected during operation was helpful for the prediction of poor CFS in patients with stage I–III CRC.

Análisis de susceptibilidad antimicrobiana y hallazgos microbiológicos: perfil en cirugía abdominal de urgencia / Microbiological findings and antimicrobial susceptibility analysis: emergency general abdominal surgery profile

Resumen Introducción La resistencia a antibióticos es un problema mundial. En los pacientes que requieren cirugía de urgencia, los antibióticos son un apoyo importante concomitante al acto quirúrgico. Objetivo Analizar los cultivos de líquido peritoneal obtenidos de pacientes operados por patología quirúrgica abdominal de urgencia. Materiales y Métodos Se realiza una cohorte prospectiva de los pacientes operados de urgencia. Se tomó cultivo de líquido peritoneal y se procesó según técnica estandarizada. Resultados Se encontró un 39,4% de cultivos positivos. E. coli fue el germen más frecuente. Destacan 5 cultivos positivos para P. aeruginosa. Existe un 25% de resistencia a ampicilina/sulbactam y 19% a quinolonas para E. coli. Conclusión La resistencia encontrada fue menor a lo reportado en la literatura, pero aún destacable. El conocimiento del perfil de bacterias y sus resistencias a antimicrobianos son importantes para las políticas hospitalarias locales de uso racional de antibióticos.

Background Antimicrobial resistance is a worldwide problem. In patients requiring emergency surgery, antibiotics are an important assistance additional to surgical intervention. Objective Analize peritoneal fluid cultures obtaines from patients who underwent emergency surgery. Methods A prospective cohort of emergency abdominal surgical patients were enrolled. Peritoneal fluid cultures were taken and processed according to standarized technique. Results A 39.4% of positive cultures was found. E. coli was the most common bacteria identified. Five positive cultures for P. aeruginosa stand out. E. coli had 25% resistance to ampicillin/sulbactam and 19% for quinolones. Conclusion Resistance found was lower than international reports, but still noteworthy. Knowledge of local bacteria profile and antimicrobial resistance is important for local antibiotic hospital policy.

Expressions and significance of IFN-γ and IL-10 mRNA in peritoneal fluid mononuclear cells of patients with endometriosis / 天津医药

Objective To investigate the expression levels and significance of interferon-γ(IFN-γ),interleukin (IL)-10 mRNA in peritoneal fluid mononuclear cells (PFMC) in patients with endometriosis (EM).Methods A total of 55 patients with EM diagnosed histopathologically after operation were enrolled as EM group,and 52 patients without EM were enrolled as control group.The PFMC were isolated,and the expression levels of IFN-γ and IL-10 mRNA were determined by TaqMan real-time reverse transcriptase-polymerase chain reaction (RT-PCR).Results It was found that IFN-γ mRNA expression level was significantly decreased in EM group compared with that in control group (P＜0.05),while IL-10 mRNA expression level was significantly increased in EM group compared with that in control group (P＜0.05).The ratio of IFN-γ/ IL-10 was significantly lower in EM group than that in control group (P＜0.05).The ratio of IFN-γIL-10 was significantly higher in patients with stage Ⅰ-Ⅱ than that in patients with stage Ⅲ-Ⅳ (P＜0.05).Conclusion The imbalance of IFN-γ/ IL-10 in PFMC is associated with the pathogenesis of EM.

Decreased Progesterone Receptor B/A Ratio in Endometrial Cells by Tumor Necrosis Factor-Alpha and Peritoneal Fluid from Patients with Endometriosis

PURPOSE: Progesterone resistance is thought to be a major factor that contributes to progression of endometriosis. However, it is not clear what causes progesterone resistance in endometriosis. This study aimed to assess whether cytokines or peritoneal fluid can affect progesterone receptor (PR) expression in endometrial cells and to verify whether PR expression is reduced in endometriosis. MATERIALS AND METHODS: The PR-B/A ratio was measured via real-time polymerase chain reaction after in vitro culture, in which endometrial cells were treated with either tumor necrosis factor-alpha (TNF-α), interleukin-1 beta, or peritoneal fluid obtained from women with advanced-stage endometriosis. Immunohistochemistry was performed to compare PR-B expression between eutopic and ectopic endometrial tissues from women with and without advanced-stage endometriosis. RESULTS: The PR-B/A ratio was significantly decreased by treatment with either TNF-α (p=0.011) or peritoneal fluid from women with advanced-stage endometriosis (p=0.027). Immunoreactivity of PR-B expression was significantly lower during the secretory phase than during the proliferative phase in endometrial tissues from control subjects (p<0.001). PR-B expression was significantly reduced in the eutopic endometrium (p=0.031) and ovarian endometrioma (p=0.036) from women with advanced-stage endometriosis compared with eutopic endometrium tissues from control subjects. CONCLUSION: Progesterone resistance in endometriosis may be caused by proinflammatory conditions in the pelvic peritoneal microenvironment.

Decreased Progesterone Receptor B/A Ratio in Endometrial Cells by Tumor Necrosis Factor-Alpha and Peritoneal Fluid from Patients with Endometriosis

PURPOSE: Progesterone resistance is thought to be a major factor that contributes to progression of endometriosis. However, it is not clear what causes progesterone resistance in endometriosis. This study aimed to assess whether cytokines or peritoneal fluid can affect progesterone receptor (PR) expression in endometrial cells and to verify whether PR expression is reduced in endometriosis. MATERIALS AND METHODS: The PR-B/A ratio was measured via real-time polymerase chain reaction after in vitro culture, in which endometrial cells were treated with either tumor necrosis factor-alpha (TNF-α), interleukin-1 beta, or peritoneal fluid obtained from women with advanced-stage endometriosis. Immunohistochemistry was performed to compare PR-B expression between eutopic and ectopic endometrial tissues from women with and without advanced-stage endometriosis. RESULTS: The PR-B/A ratio was significantly decreased by treatment with either TNF-α (p=0.011) or peritoneal fluid from women with advanced-stage endometriosis (p=0.027). Immunoreactivity of PR-B expression was significantly lower during the secretory phase than during the proliferative phase in endometrial tissues from control subjects (p<0.001). PR-B expression was significantly reduced in the eutopic endometrium (p=0.031) and ovarian endometrioma (p=0.036) from women with advanced-stage endometriosis compared with eutopic endometrium tissues from control subjects. CONCLUSION: Progesterone resistance in endometriosis may be caused by proinflammatory conditions in the pelvic peritoneal microenvironment.

Evaluation of semi-automated cells counting in peritoneal fluid / Avaliação de semiautomação para contagem de células em líquido peritoneal

ABSTRACTIntroduction:Currently, the cytological analysis of biological fluids, such as peritoneal fluid, is performed by manually cells counting in Fuchs-Rosenthal chamber. However, this method has a number of limitations. Because of these limitations, automatic counters have been evaluated for cell counting in this type of sample in order to make it faster and more reliable test.Objective:The aim of this study is to compare the manual and semi-automated leukocytes and erythrocytes counting in peritoneal fluid.Materials and methods:The samples were analyzed manually and using the CountessTM(Invitrogen).Results:The results showed that although there is a correlation between the two counting methods, the correlation is relatively low, for both leukocytes and erythrocytes analysis.Conclusion:The results suggest that peritoneal fluid should continue to be analyzed in Fuchs-Rosenthal chamber. However, further studies should be conducted with a greater number of samples to investigate the possibility of using automated cells counting in serous fluids and, thus, provide greater speed and quality of results.

RESUMOIntrodução:Atualmente, a análise citológica de líquidos biológicos, como líquido peritoneal, é realizada por meio da contagem manual de células, em câmara de Fuchs-Rosenthal. Porém, esse método apresenta uma série de limitações. Com isso, contadores automáticos têm sido avaliados para a contagem de células nesse tipo de amostra a fim de tornar esse exame mais rápido e confiável.Objetivo:Comparar a contagem manual e semiautomatizada de leucócitos e eritrócitos em líquido peritoneal.Materiais e métodos:As amostras foram analisadas manualmente e no contador de células CountessTM (Invitrogen).Resultados:Os resultados mostraram que apesar de existir correlação entre os dois métodos de contagem, essa correlação é relativamente fraca, tanto para análise de leucócitos como para de eritrócitos.Conclusão:Esses resultados sugerem que o líquido peritoneal deve continuar a ser analisado em câmara de Fuchs-Rosenthal, contudo novos estudos devem ser realizados, com maior número de amostras, para investigar a possibilidade do uso de automação na contagem de células em líquidos serosos e, assim, proporcionar maior agilidade e qualidade no resultado.

Influência do parasitismo intestinal sobre os parâmetros hematológicos e de líquido peritoneal em equinos de tração / Influence of intestinal parasitism on peritoneal fluid and hematological parameters in horses used for traction

O objetivo do presente estudo foi avaliar a influência do parasitismo intestinal crônico sobre parâmetros hematológicos e de líquido peritoneal por meio da comparação dessas características em equinos naturalmente parasitados e após administração de anti-helmíntico. Utilizaram-se 21 cavalos de tração urbana, entre dois e 19 anos, sem raça definida e com resultado de exame parasitológico superior a 300 ovos por grama de fezes. Foi realizada avaliação física e coleta de fezes, de líquido peritoneal e de sangue em dois momentos do experimento (D0 e D15), sendo efetuado tratamento antiparasitário no D0. No fluido peritoneal foram avaliadas características físicas, bioquímicas, bem como contagem de células nucleadas (CTCN) e diferenciação celular. No sangue foram determinados valores eritrocitários, leucocitários, proteínas plasmáticas totais, glicose e fibrinogênio plasmáticos, além de fosfatase alcalina (FA) sérica. A análise dos parâmetros avaliados não demonstrou diferença significativa entre animais parasitados e após administração de anti-helmíntico, exceto para valores de CTCN, contagem de neutrófilos segmentados e grau de turbidez do líquido peritoneal. As médias se mantiveram dentro dos intervalos de referência, com exceção da CTCN do líquido peritoneal no D0. No líquido peritoneal, houve predomínio de neutrófilos segmentados, seguidos por macrófagos, linfócitos e eosinófilos em ambos os momentos de avaliação. Observaram-se tendência do quadro eritrocitário em manter-se próximo aos limites inferiores e leve leucocitose no D0. A infecção parasitária nos animais estudados foi predominantemente moderada, o que oferece poucos riscos clínicos. Nessas condições, pode-se afirmar que a CTCN, a contagem absoluta de neutrófilos segmentados e o grau de turbidez do líquido peritoneal são influenciados e podem ser considerados ferramentas diagnósticas e prognósticas úteis nas parasitoses intestinais crônicas.(AU)

The aim of this study was to evaluate the influence of chronic intestinal parasitism on hematological parameters and peritoneal fluid. It was done by comparing these features in horses used for traction naturally parasitized and after the administration of anthelmintic. Twenty-one horses, between two and nineteen years of age, of mixed breed and with results of parasitological examination of more than 300 eggs per gram of feces were studied. Physical assessment and samples of feces were conducted, as well as blood and peritoneal fluid in the two phases of the experiment (D0 and D15). Antiparasitic treatment in D0 has also been done. The peritoneal fluid was evaluated for physical and biochemical features, and also total count of nucleated cells (TCNC) and cell differentiation. The blood was determined for erythrocyte, leukocyte, plasma total protein, glucose and plasma fibrinogen, and alkaline phosphatase (ALP) levels. The analysis of these parameters showed no significant difference between parasitized animals and after administration of anthelmintic except for TCNC values, segmented neutrophil count and degree of turbidity in peritoneal fluid. The averages remained within the reference ranges, except the TCNC in the peritoneal fluid in D0. In the peritoneal fluid there was a predominance of segmented neutrophils, followed by macrophages, lymphocytes and eosinophils in both time points. A trend was observed in erythrocyte frame to keep close to the lower limits and mild leukocytosis in D0. Parasitic infection of the animals studied was predominantly moderate, which offered minimal clinical risks. After that, it can be affirmed that the TCNC, absolute segmented neutrophil count and targeted degree of turbidity in peritoneal fluid are influenced and can be considered useful diagnostic and prognostic tools in chronic intestinal parasitism.(AU)

Evaluation of the Usefulness of MacConkey Agar and Colistin-Nalidixic Acid Blood Agar for Body Fluids, Peritoneal Fluid, and Wound/Abscess Specimens / 대한임상미생물학회지

BACKGROUND: Most clinical microbiology laboratories in Korea have difficulty in following the recommendations of the clinical procedure handbook for culture of body fluid and wound/abscess specimens. We evaluated the usefulness of MacConkey (MAC) and colistin-nalidixic acid blood agar (CNA) for the isolation of pathogens from these specimens. METHODS: A total of 1,508 clinical specimens [144 peritoneal fluid, 241 body fluids (19 bile, 70 joint fluid, 6 pericardial fluid, 104 pleural fluid, and other fluids in 42 cases) and 1,123 wound/abscess] were inoculated onto basic media [Blood agar plate (BAP), chocolate agar or BAP with streaking of Staphylococcus aureus] and simultaneously inoculated onto MAC and CNA. The pathogens isolated by basic media and by additional use of MAC and/or CNA were compared. RESULTS: With basic media, 885 isolates from 588 specimens were detected, and by additional use of MAC and CNA, an additional 27 isolates from 24 specimens and an additional 128 isolates from 112 specimens were isolated, respectively. Compared to the basic media, by adding MAC, an additional 233.3%, 38.5% and 4.5% of gram-negative bacteria were isolated from peritoneal fluids, body fluid and wound/abscess, respectively, and by adding CNA, an additional 106.7%, 45.0%, and 20.7% of gram-positive bacteria/ yeast were isolated, respectively. The isolates detected by additional use of MAC were mainly Enterobacteriaceae (77.0%), and those detected by CNA were S. aureus (21.1%), Coagulase-negative Staphylococcus spp. (20.3%), Enterococcus spp. (16.4%), Streptococcus spp. (10.2%) and yeasts (16.4%). CONCLUSION: For peritoneal fluid and body fluid specimens, additional use of MAC plus CNA seems necessary for detection of pathogens. For wound/abscess, additional use of CNA will be cost effective.

Establishment of peritoneal liquid electrophoretogram from healthy horses and horses submitted to experimentally induced intestinal obstruction / Estabelecimento do eletroforetograma do líquido peritoneal de equinos hígidos e daqueles submetidos à obstrução intestinal experimental

The initial inflammatory stages of the colic syndrome include changes known as acute phase response. The aim of this study was to contribute with the establishment of reference values concerning the electrophoretogram of peritoneal liquid from healthy horses and horses submitted to experimentally induced intestinal obstruction. Twenty-one horses were allotted in four groups: duodenal obstruction (DG), ileum obstruction (IG), left-dorsal colon obstruction (MG), and control group (CG). Peritoneal liquid was sampled before obtruction (T0), with 3 hours of obstruction (T3) and 6, 30, 102 and 174 hours after desobstructing (T6, T30, T102 and T174, respectively). Total protein levels were determined by the biuret method and protein fractions were obtained by SDS-PAGE electrophoresis. The acute phase proteins (APP) identified were Immunoglobulin-A, ceruloplasmin, transferrin, albumin, α1-antitrypsin, heavy and light chains of immunoglobulin-G, haptoglobin, α1-acid glycoprotein and a still unnamed protein, which was called P24. There was no difference (P>0.3) in protein levels among groups, although a significant difference (P>0.05) was observed between distinct experimental moments in each group evidencing a higher response of the APP in the obstructed groups. The APP fractioning of the peritoneal liquid was standardized to establish a standard curve for healthy equines and those submitted to induced intestinal obstruction. Moreover, it was verified that the SDS-PAGE electrophoresis was sensitive and effective to help diagnose abdominal inflammatory processes...

Na cólica equina, os estágios iniciais da inflamação incluem alterações denominadas resposta de fase aguda. O objetivo deste estudo foi contribuir para o estabelecimento de valores de referência do proteinograma do líquido peritoneal de equinos hígidos e daqueles submetidos à obstrução intestinal experimental. Vinte e um animais foram distribuídos nos grupos: obstrução de duodeno (GD), íleo (GI), cólon dorsal esquerdo (GM) e controle instrumentado (GC). As colheitas das amostras de líquido peritoneal foram realizadas antes (T0), durante as obstruções (T3) e após as desobstruções (T6, T30, T102 e T174 horas). A proteína total foi determinada pelo método do biureto, e as frações proteicas obtidas por eletroforese em SDS-PAGE. Identificaram-se as proteínas de fase aguda (PFA): IgA, ceruloplasmina, transferrina, albumina, α1-antitripsina, cadeias pesada e leve de imunoglobulina-G, haptoglobina, alfa-1-glicoproteína ácida e uma proteína nominalmente não identificada, que foi chamada P24. Não houve diferença (P>0.3) nas concentrações proteicas entre os grupos, somente entre tempos dentro de cada grupo (P>0.05), evidenciando uma resposta maior das PFA dos grupos obstruídos. O fracionamento eletroforético das PFA, presentes no líquido peritoneal, foi padronizado de modo a estabelecer a curva-padrão para equinos hígidos e para aqueles submetidos à obstrução intestinal; ademais, verificou-se que o referido fracionamento proteico mostrou-se sensível e eficaz no auxílio ao diagnóstico de processos inflamatórios abdominais...

Proteinograma sérico e do líquido peritoneal de equinos hígidos e daqueles submetidos à obstrução intestinal experimental / Serum and peritoneal protein profile of healthy horses and horses subjected to experimental intestinal obstruction

Avaliou-se a resposta de fase aguda através da concentração das proteínas de fase aguda (PFA) no soro sanguíneo e no líquido peritoneal de vinte e um equinos, hígidos e submetidos à obstrução intestinal experimental, distribuídos em quatro grupos: obstrução de duodeno - GD (n=6), íleo - GI (n=6), cólon dorsal esquerdo - GM (n=6) e controle instrumentado - GC (n=3). Foram colhidas amostras de sangue e líquido peritoneal e, após centrifugação e fracionamento, as proteínas de fase aguda foram separadas por eletroforese em SDS-PAGE. Identificaram-se as proteínas IgA, ceruloplasmina, transferrina, albumina, IgG, haptoglobina, α1-glicoproteína ácida e P24, no soro e no líquido peritoneal. Houve aumento nas concentrações sérica e peritoneal de todas as PFA, sendo mais evidente no líquido peritoneal e nos animais obstruídos. O fracionamento eletroforético das PFA no líquido peritoneal é mais eficaz no diagnóstico de processos inflamatórios abdominais, quando comparado ao sérico.

It was evaluated the acute phase response over the concentration of acute phase proteins (APP) in serum and in peritoneal fluid of twenty-one healthy equines submitted to experimental intestinal obstruction distributed in groups: duodenal obstruction (GD), ileum obstruction (GI), left-dorsal colon (GM), and instrumentalized control (GC). Blood and peritoneal liquid samples were collected, after centrifugation and fractioning and the acute phase proteins were separated by electrophoresis in SDS-PAGE. The acute phase proteins (APP) identified were IgA, ceruloplasmin, transferrin, albumin, IgG, haptoglobin, α1-acid glycoprotein and P24, in the serum and liquid peritoneal. There was anincrease in the serum and peritoneal proteins concentrations evidencing a higher response of the APP from the peritoneal fluid and of the obstructed groups. The electrophoretic separation of the proteins included in the peritoneal liquid is more effective when compared to the serum concentration in the diagnosis of abdominal inflammatory processes.

Cytological Study of Effusions.

Background: Effusion fluid analysis plays an important role in clinical medicine. Clinicians rely on the reports of effusion fluids and use them as complement to their clinical assessment for the diagnosis and management. Aim: To study the incidence of neoplastic and non neoplastic effusions. Objectives: i) To study the gross and microscopic features of effusions; ii) To study the pattern of effusions in various neoplastic and non neoplastic conditions. Material and Method: 550 specimens of pleural, peritoneal and pericardial fluid were studied. Fluid samples were centrifuged for five minutes at 2000 rpm and smears prepared from deposit were stained by Haematoxylin and Eosin (H and E), Giemsa and Papanicolaou stains (Pap). Result: Out of 550, 315 were pleural effusions, 234 peritoneal and one was pericardial. Out of total 315 cases of pleural effusions, 297 were non neoplastic and 18 were neoplastic effusion. Out of total 234 peritoneal effusions 214 were non neoplastic and 20 neoplatic. Commonest malignancy in pleural and peritoneal fluid was adenocarcinoma. Conclusion: Pleural effusion was the commonest fluid in this study. Exudates were predominant in pleural effusion and transudates were predominant in peritoneal effusion. Common causes of exudates in pleural effusion were tuberculosis (TB), pneumonia and malignancy. Common causes of transudates in peritoneal effusion were liver cirrhosis and congestive cardiac failure (CCF). Adenocarcinoma was the commonest malignancy in both pleural and peritoneal effusion (30 cases).

Cancer cells with p53 deletion detected by fluorescent in situ hybridization in peritoneal drainage fluid is correlated with early peritoneal seeding in resectable pancreatic cancer / 대한외과학회지

PURPOSE: Free tumor cells in peritoneal fluid in patients with pancreatic cancer may have prognostic significance but there are few reports on methods for the effective detection of free tumor cells. The aims of this study were to identify free cancer cells in peritoneal fluid with fluorescent in situ hybridization (FISH) technique and to investigate its prognostic significance. METHODS: Twenty-eight patients with resectable pancreatic cancer who underwent surgical resection were included. Peritoneal washing and peritoneal drainage fluid were examined by FISH for p53 deletion. RESULTS: Among the study subjects, the R0 resection rate was 75%. None of the patients had positive cytology with Papanicolaou's method. p53 deletion was detected in 9 peritoneal washings (32.1%) and in 5 peritoneal drainage fluids (17.9%). After a median of 18 months of follow-up, 25 patients (89.3%) experienced recurrence and 14 patients (50.0%) had peritoneal seeding. Patients with p53 deletion detected in the peritoneal drainage fluid had positive radial margin (60.0% vs. 17.4%, P = 0.046) more frequently and a lower peritoneal metastasis free survival (median, 11.1 months vs. 30.3 months; P = 0.030). Curative resection (P < 0.001) and p53 deletion in peritoneal drainage fluid (P = 0.030) were independent risk factors of peritoneal metastasis free survival after multivariate analysis. CONCLUSION: FISH technique detects free cancer cells with higher sensitivity compared to Papanicolaou's method. p53 deletion detected in peritoneal drainage fluid is correlated with positive radial resection margin and results in early peritoneal seeding. Patients with p53 deletion in peritoneal drainage fluid need more aggressive adjuvant treatment.

Peritoneal fluid changes in horses subjected to small colon distension / Alterações no líquido peritoneal de equinos submetidos a distensão do cólon menor

Intestinal devitalization in cases of small colon obstruction may be difficult to detect based only in clinical signs. The purpose was to serially evaluate blood and peritoneal fluid of horses subjected to small colon distension. Seventeen adult horses were allotted in three groups. In the small colon-distended group (DG, n=7) a surgically-implanted latex balloon was inflated to promote intraluminal small colon distension. In the shamoperated group (SG, n=5), the balloon was implanted but not inflated, and no surgery was done in the control group (CG, n=5). Blood and peritoneal fluid were sampled before and after (6 samples with a 30-minute interval) intestinal obstruction for cytological and biochemical analyses. No significant changes in clinical signs occurred within groups or across time during the experimental period. There were no statistical differences among SG and SG groups in hematologic and blood chemistry variables. Although total protein concentration and lactate dehydrogenase (LDH) activity in peritoneal fluid remained most of the time within reference values during the experimental period in all groups, increases from baseline values were detected in SG and DG groups. Such increases occurred earlier, progressively and with greater magnitude in the DG when compared with the SG (P<0.05). Increases from baselines values were also observed in total nucleated cells and neutrophils counts in the DG (P<0.05). In conclusion, distension of the equine small colon induced progressive subtle increases in total protein and LDH concentrations in the peritoneal fluid during the first hours. Serial evaluation of these variables in peritoneal fluid may be useful for early detection of intestinal devitalization in clinical cases of equine small colon obstruction.

A desvitalização do cólon menor em equinos pode ser difícil de ser detectada baseando-se apenas em sinais clínicos. O objetivo foi realizar uma avaliação seriada do líquido peritoneal de equinos submetidos à distensão do cólon menor. Dezessete cavalos adultos foram divididos aleatoriamente em três grupos. No grupo distendido (DG, n=7) um balão implantado cirurgicamente foi inflado para promover distensão do cólon menor. No grupo instrumentado (SG, n=5) o balão foi implantado, mas sem promover distensão e no grupo controle (CG, n=5) não houve anestesia ou cirurgia. Sangue e fluido peritoneal foram colhidos antes e durante 180 minutos após a cirurgia para análises citológicas e bioquímicas. Nenhuma interação significativa ocorreu entre grupos e tempos nas variáveis clínicas e hematológicas. Apesar dos valores de proteínas totais e da atividade da lactato desidrogenase (LDH) permanecerem dentro da normalidade durante quase todo o experimento, aumentos em relação aos valores basais ocorreram nos grupos SG e DG. Contudo, tais aumentos foram precoces, progressivos e em maior magnitude em DG quando comparados ao SG, mostrando que a distensão promoveu alterações significativas nessas variáveis (P<0.05). Aumentos em relação aos valores basais também ocorreram nas contagens de células totais nucleadas e neutrófilos (P<0.05). Em conclusão, a distensão experimental do cólon menor promove, nas primeiras horas, alterações subliminares progressivas nas concentrações de proteínas totais e na atividade de LDH no líquido peritoneal. Os resultados indicam que a avaliação seriada do liquido peritoneal pode ser útil para detectar desvitalização intestinal em casos clínicos de obstrução do cólon menor equino.

Vaccination with Trypanosoma rangeli modulates the profiles of immunoglobulins and IL-6 at local and systemic levels in the early phase of Trypanosoma cruzi experimental infection

In America, there are two species of Trypanosoma that can infect humans: Trypanosoma cruzi, which is responsible for Chagas disease and Trypanosoma rangeli, which is not pathogenic. We have developed a model of vaccination in mice with T. rangeli epimastigotes that protects against T. cruzi infection. The goal of this work was to study the pattern of specific immunoglobulins in the peritoneum (the site of infection) and in the sera of mice immunized with T. rangeli before and after challenge with T. cruzi. Additionally, we studied the effects triggered by antigen-antibodies binding and the levels of key cytokines involved in the humoral response, such as IL-4, IL-5 and IL-6. The immunization triggered the production of antibodies reactive with T. cruzi in peritoneal fluid (PF) and in serum, mainly IgG1 and, to a lesser magnitude, IgG2. Only immunized mice developed specific IgG3 antibodies in their peritoneal cavities. Antibodies were able to bind to the surface of the parasites and agglutinate them. Among the cytokines studied, IL-6 was elevated in PF during early infection, with higher levels in non-immunized-infected mice. The results indicate that T. rangeli vaccination against T. cruzi infection triggers a high production of specific IgG isotypes in PF and sera before infection and modulates the levels of IL-6 in PF in the early periods of infection.