Changes in tissue perfusion parameters in dogs with severe sepsis/septic shock in response to goal-directed hemodynamic optimization at admission to ICU and the relation to outcome.

OBJECTIVE: To evaluate the changes in tissue perfusion parameters in dogs with severe sepsis/septic shock in response to goal-directed hemodynamic optimization in the ICU and their relation to outcome. DESIGN: Prospective observational study. SETTING: ICU of a veterinary university medical center. ANIMALS: Thirty dogs with severe sepsis or septic shock caused by pyometra who underwent surgery and were admitted to the ICU. MEASUREMENTS AND MAIN RESULTS: Severe sepsis was defined as the presence of sepsis and sepsis-induced dysfunction of one or more organs. Septic shock was defined as the presence of severe sepsis plus hypotension not reversed with fluid resuscitation. After the presumptive diagnosis of sepsis secondary to pyometra, blood samples were collected and clinical findings were recorded. Volume resuscitation with 0.9% saline solution and antimicrobial therapy were initiated. Following abdominal ultrasonography and confirmation of increased uterine volume, dogs underwent corrective surgery. After surgery, the animals were admitted to the ICU, where resuscitation was guided by the clinical parameters, central venous oxygen saturation (ScvO(2)), lactate, and base deficit. Between survivors and nonsurvivors it was observed that the ScvO(2), lactate, and base deficit on ICU admission were each related independently to death (P = 0.001, P = 0.030, and P < 0.001, respectively). ScvO(2) and base deficit were found to be the best discriminators between survivors and nonsurvivors as assessed via receiver operator characteristic curve analysis. CONCLUSION: Our study suggests that ScvO(2) and base deficit are useful in predicting the prognosis of dogs with severe sepsis and septic shock; animals with a higher ScvO(2) and lower base deficit at admission to the ICU have a lower probability of death.

Identificação das bactérias envolvidas na sepse grave de fêmeas caninas com piometra submetidas a ováriohisterectomia terapêutica / Identification of the microorganisms involved in severe sepsis caused by pyometra in bitches undergoing ovariohysterectomy

A piometra é uma afecção reprodutiva comum que acomete fêmeas caninas, podendo se agravar e progredir para o quadro de sepse grave e choque séptico. A precocidade da instituição da antibioticoterapia é determinante para um melhor prognóstico. O objetivo deste estudo foi avaliar os principais microrganismos envolvidos nos casos de sepse grave em cadelas acometidas por piometra e submetidas à ovário-histerectomia terapêutica, por meio de realização de hemocultura e cultura da secreção uterina e antibiograma. Foram avaliadas 33 fêmeas caninas e o principal agente envolvido com a sepse grave secundária à piometra foi a Escherichia coli, identificada em 57,57% dos casos. Também foram identificados Staphylococcus sp., com incidência de 9,09%, Citrobacter koseri, Enterobacter cloacae, Enterobacter faecalis, Eduardsiella sp., Klebsiella pneumoniae e Streptococcus sp., com 3,03% de frequência cada. Após a realização do antibiograma pelo método de difusão, os antimicrobianos que apresentaram maior eficácia contra as cepas de Escherichia coli foram a gentamicina, a enrofloxacina, a cefalexina e a associação de amoxicilina com ácido clavulânico, nesta ordem. A cultura da secreção uterina foi mais sensível que a hemocultura para identificação do agente microbiano (p<0,0001). A identificação bacteriana é útil para direcionar a antibioticoterapia empírica mais específica, de acordo com o perfil de sensibilidade, minimizando assim o desenvolvimento de resistência, o custo do tratamento e o risco de reações adversas aos antimicrobianos utilizados.

Pyometra is a common reproductive disorder that affects female dogs. It can represent a serious clinical entity and progress to severe sepsis and septic shock. The premature antibiotic therapy is crucial for a better prognosis. The aim of this study was to identify the most frequent microorganisms involved in the context of pyometra and severe sepsis in animals undergoing ovariohysterectomy, by blood and uterine secretion culture and antibiogram. The tests were conducted in 33 female dogs with pyometra. The most frequent recovered bacteria were Escherichia coli in 57.57%. Staphylococcus sp. was also isolated, in 9.09%. Citrobacter koseri, Enterobacter cloacae, Enterobacter faecalis, Eduardsiella sp., Klebsiella pneumoniae e Streptococcus sp. were also found with a 3.03% frequency each. After diffusion antibiotic test, the gentamicin was the most effective drug to E. coli, followed by enrofloxacin, cephalexin and the combination of amoxicillin and clavulanic acid. Uterine secretions cultures were more sensitive than blood culture to identify the bacterial (p<0.0001). The bacterial identification followed by an antibiogram allows to choose a better therapy in the presented disease in dogs.

Avaliação ultrassonográfica de linfonodos na pesquisa de metástases de neoplasia mamária em cadelas / Ultrasonographic evaluation of lymph nodes for metastasis research of canine mammary tumor

A avaliação de linfonodo, parte importante do estadiamento das neoplasias mamárias em cadelas, pode auxiliar no estabelecimento do prognóstico e na escolha da conduta terapêutica. A ultrassonografia em modo B possibilita avaliação de tamanho, contorno, borda, forma, arquitetura, ecotextura e ecogenicidade do parênquima dos linfonodos e, em modo Doppler, da quantidade e distribuição dos seus vasos internos. Este trabalho visou identificar as características ultrassonográficas mais importantes utilizadas para classificar os linfonodos em metastáticos e não-metastáticos, estabelecer elementos de confiabilidade do ultrassom como ferramenta para diferenciar linfonodos metastáticos de não-metastáticos e estabelecer procedimentos de reprodução deste exame. Foram examinados 67 linfonodos inguinais superficiais de 30 cadelas com tumor mamário e cada linfonodo foi classificado como metastático ou não-metastático. A impressão diagnóstica ultrassonográfica foi associada aos resultados do exame histopato lógico dos linfonodos obtendo-se taxa de concordância de 92,5 por cento, índice de sensibilidade de 94,1 por cento, índice de especificidade de 92 por cento, valor preditivo positivo de 0,8 e valor preditivo negativo de 0,9787. Características ultrassonográficas que classificaram um linfonodo como metastático ou como não-metastático foram listadas. O exame ultrassonográfico dos linfonodos regionais constitui importante ferramenta na detecção de metástase e sugere-se incluí-lo como rotina do estadiamento de neoplasias mamárias em cadelas.

Evaluation of lymph nodes is part of the staging of the mammary tumors and helps to establish a prognosis and therapy. B-mode ultrasonography was used to evaluate size, edge, nodal borders, shape, architecture, echotexture and echogenicity of lymph nodes and Doppler ultrasonography to evaluate the quantity and distribution of the vessels. The aims of this work were to identify which ultrasound features can be used to classify a lymph node as metastatic or non-metastatic; to establish its accuracy as a useful tool to this differentiation, and to establish procedures that permit the reproducibility of the results. Sixty seven lymph nodes of 30 female dogs with mammary tumors were examined by ultrasound and classified as metastatic or non-metastatic. The ultrasonographic diagnostic impression was confronted with the histopathologic results. The ultrasonographic classification of the lymph nodes was correct in 92.5 percent of the cases, with sensibility of 94.1 percent, specificity of 92 percent, positive predictive value of 0.8, and negative predictive value of 0.9787.The characteristics of the nodes to classify them as metastatic or non metastatic were registered. Ultrasound imaging of regional lymph nodes is important to detect metastases and it is suggested that it becomes a regular procedure in the staging of canine mammary tumors.

Avaliação da terapia com fluido no período perioperatório da ovariossalpingohisterectomia, em cadelas com piometra e insuficiência renal aguda / Avaliation of perioperative conservative treatment in pyometra ovariohysterectomy bitches of in acute renal failure

A insuficiência renal aguda foi avaliada em 351 cadelas com piometra, selecionando-se 132 pelos critérios: creatinina > 2,4 mg/dL e/ou ureia > 80 mg/dL e/ou aumentos relativos de 100% de creatinina e/ou ureia séricas, no diagnóstico, 24 horas após a ovariossalpingohisterectomia e na retirada dos pontos. Foi administrado Ringer lactato de sódio até 90 mL/kg/h, durante quatro horas, ou três dias. Os parâmetros avaliados antes e após cada terapia foram: peso, pressão arterial sistólica (PAS), frequência cardíaca, função renal, hemogasimetria arterial, bioquímica sérica e urinária, unidade de remoção de ureia, fração de excreção de eletrólitos e clearance de creatinina. A análise univariada detectou diferença significante (p < 0,05) para: PAS, pH urinário, peso corpóreo, ureia e creatinina séricas, fração de excreção de eletrólitos e clearance da creatinina. Comparadas as sobreviventes (79,6%) com as não sobreviventes (20,4%), respectivamente, detectou-se creatinina sérica de 2,38 ± 1,33 mg/dL (média ± desvio padrão) e mediana 2,2 mg/dL e de 5,54 ± 3,0 mg/dL (média ± desvio padrão) e mediana 4,4 mg/dl, com diferença significante (p < 0,0001). Quanto ao clearance da creatinina endógena nas sobreviventes, 1,82 ± 1,74 mL/kg/min (média ± desvio padrão) e mediana 1,34 mg/dL e as não sobreviventes 0,36 ± 0,38 mg/dL (média ± desvio padrão) e mediana 0,23 mg/dL, com diferença significativa (p < 0,0001). Conclui-se que o critério de inclusão pode auxiliar na seleção dos animais em estado grave e o clearance da creatinina, considerado relevante quando observados valores > 1 mL/kg/min; negativo com valores < 1 mL/kg/min e indicador de falência renal grave e ou morte para valores < 0,5 ml/kg/min.

Acute renal failure (ARF) was evaluated prospectively in 351 female dogs with piometra, 132 animals were selected for this study, fulfilling the following criteria, measured in three moments (immediately after the diagnosis, 24 hours after the hysterectomy and when the suture was removed): creatinine > 2.4 mg/dL and/or urea > 80 mg/dL and/or relative increases of 100% in comparison with values previously obtained. The animals selected for the prospective study received conservative treatment consisting in lactated Ringer solution, with the maximum dose of 90 mL/kg/hr, during four hours. The factors evaluated before and after the therapy were: weight, systolic arterial pressure, heart rate, renal function, arterial hemogasometry, serum and urinary biochemistry and the endogenous renal creatinine clearance. The evaluated parameters showed significant statistical differences (p < 0.05) when companing: PAS (systolic arterial pressure), urinary pH, wheigt, serum urea and creatinine, excretion electrolyte rate and creatinine clearance. Howerer, the survining animals (79.6%) that received the treatment described above showed decresead levels of serum creatinine- 2,38 ± 1,33mg/dL and median 2.2 mg/dL – when compared to the non-survining animals (20.4%) that received the same treatment – 5.54 ± 3.0mg/dL and median 4.4mg/dL – (p < 0.0001). When comparing the endogenous creatinine clearance, the survining animals had higher values – 1.82 ± 1.74mL/kg/min and median 1.34 mL/kg/min – than the non-survining animals – 0.36 ± 0.38 mL/kg/min and median 0.23 mL/kg/min – (p < 0.0001). Concluding: the criteria used to include the animals in this study can help to select the severely affected animals. Values higher than 1mL/kg/min are considered as a good prognosis while values lower than 1mL/kg/min are considered as a restricted prognosis. Values lower than 0.5 mL/kg/min are considered as strong predictors of severe renal failure or death.

Effects of gonadotropin-exposed medium with high concentrations of progesterone and estradiol-17beta on in vitro maturation of canine oocytes.

During the process of maturation in the oviduct, canine oocytes in the germinal vesicle stage are exposed to decreasing levels of estradiol-17beta and increasing levels of progesterone. However, hormone concentrations in the microenvironments in which they act are higher than serum concentrations. Therefore, the aim of the present study was to compare the meiotic competence of canine oocytes harvested from anestrous bitches in culture medium containing high concentrations (20 microg ml(-1)) of estradiol-17beta and/or progesterone in association to gonadotropins (luteinizing hormone and follicle-stimulating hormone) using three different maturation periods (48, 72, and 96 h). Oocytes were cultured in tissue culture medium (TCM-199) and arranged in four experimental groups: group control, group E2 (estradiol-17beta), group P4 (progesterone), and group E2 + P4. Regardless of the maturation period, groups P4 and E2 + P4 presented statistically higher rate of germinal vesicle breakdown oocytes compared to the group control and group E2. There were no significant differences among groups on germinal vesicle, metaphase I, metaphase II, and degenerated or unidentifiable oocytes rates. The mean percentage of metaphase II oocytes was higher at 96 h when compared to 72 h. Results of the present research indicate no influence of estradiol-17beta supplementation, unless in association with progesterone. There is an evidence of the positive effect of progesterone on germinal vesicle breakdown. Results also showed that extended periods of in vitro maturation affect positively maturation rates to metaphase II of low competent oocytes harvested from anestrous bitches, independent of the maturation media. In conclusion, high concentrations of steroids, especially progesterone, have positive effect on in vitro oocyte maturation when the oocytes are derived from the anestrous status.

Microbiological and histopathological aspects of canine pyometra / Aspectos microbiológicos e histopatológicos da piometria canina

As pyometra is recognized as one of the main causes of disease and death in the bitch the purposes of this study were to evaluate microbiological and histopathological aspects of canine pyometra and to research the virulence factors of the E. coli isolates identifying possible risks to human health. The microbiological isolation from the intrauterine contents of 100 dogs with pyometra was carried out and the virulence factors in the E. coli strains were identified using PCR method. This study also consisted of the counting of microorganisms colonies forming units in samples of intrauterine content, tests of antimicrobial susceptibility of the E. coli isolates and the histological examination of the uterus. E. coli was the most prevalent microorganism isolated (76.6 percent) and 120 strains (79.5 percent) were positive for sfa, 86 (56.9 percent) were positive for cnf, 87 (57.6 percent) were positive for pap, 52 (34.4 percent) were positive for hly, 51 (33.8 percent) were positive for iuc and 5 (3.3 percent) were positive for afa genes. One observed more sensitivity of E. coli to norfloxacin, polimixin B, sulphazotrin, chloranfenicol and enrofloxacin. In 42 percent of the samples of uterine walls where microorganisms were isolated, the sizes of the areas of the inflammatory responses corresponded to 39-56 percent. Virulence factors were identified in 98.0 percent of the strains evaluated, demonstrating a high frequency of potentially pathogenic E. coli. It must be considered that dogs are animals that are living in close proximity to man for thousands of years and have an important role in the transmission of E. coli to other animals and to man.

A piometra é uma enfermidade da cadela adulta, sendo a doença reconhecida como uma das causas mais comuns de morte desta espécie animal. Os objetivos deste trabalho foram a avaliação de aspectos microbiológicos e histopatológicos da piometra canina e pesquisa de fatores de virulência de E. coli, identificando possíveis riscos para a saúde humana. Foi realizado o exame microbiológico de conteúdo intra-uterino de 100 cadelas com piometra bem como a contagem de unidades formadoras de colônias de microrganismos nestas amostras, testes de susceptibilidade "in vitro" aos antimicrobianos de E. coli e pesquisa de fatores de virulência nestas estirpes, e exame histopatológico de amostras de útero. Escherichia coli foi o microrganismo mais freqüentemente isolado (76,6 por cento), sendo que 120 estirpes (79,5 por cento) foram positivas para os genes sfa, 86 (56,9 por cento) foram positivas para cnf, 87 (57,6 por cento) foram positivas para pap, 52 (34,4 por cento) foram positivas para hly, 51 (33,8 por cento) foram positivas para iuc e 5 (3,3 por cento) foram positivas para afa. Observou-se maior sensibilidade de E.coli à norfloxacina, polimixina B, sulfazotrim, cloranfenicol e enrofloxacina. Em 42 por cento das amostras de parede uterina nas quais foram isolados microrganismos, os tamanhos das áreas de processo inflamatório corresponderam a 39-56 por cento. Foram identificados fatores de virulência em 98,0 por cento das estirpes de Escherichia coli avaliadas, demonstrando uma alta freqüência de E. coli potencialmente patogênica. Deve-se considerar que os cães são animais que vivem em proximidade aos homens há milhares de anos e possuem um papel importante na transmissão de E. coli aos outros animais e também ao homem.

Microbiological and histopathological aspects of canine pyometra.

As pyometra is recognized as one of the main causes of disease and death in the bitch the purposes of this study were to evaluate microbiological and histopathological aspects of canine pyometra and to research the virulence factors of the E. coli isolates identifying possible risks to human health. The microbiological isolation from the intrauterine contents of 100 dogs with pyometra was carried out and the virulence factors in the E. coli strains were identified using PCR method. This study also consisted of the counting of microorganisms colonies forming units in samples of intrauterine content, tests of antimicrobial susceptibility of the E. coli isolates and the histological examination of the uterus. E. coli was the most prevalent microorganism isolated (76.6%) and 120 strains (79.5%) were positive for sfa, 86 (56.9%) were positive for cnf, 87 (57.6%) were positive for pap, 52 (34.4%) were positive for hly, 51 (33.8%) were positive for iuc and 5 (3.3%) were positive for afa genes. One observed more sensitivity of E. coli to norfloxacin, polimixin B, sulphazotrin, chloranfenicol and enrofloxacin. In 42% of the samples of uterine walls where microorganisms were isolated, the sizes of the areas of the inflammatory responses corresponded to 39-56%. Virulence factors were identified in 98.0% of the strains evaluated, demonstrating a high frequency of potentially pathogenic E. coli. It must be considered that dogs are animals that are living in close proximity to man for thousands of years and have an important role in the transmission of E. coli to other animals and to man.

Immunohistochemical determination of estrogen receptor-alpha in vaginal and tumor tissues of healthy and TVT-affected bitches and their relation to serum concentrations of estradiol-17beta and progesterone.

One of the most frequent canine neoplasms is the transmissible venereal tumor (TVT), which affects the male and the female genital tract. The objective of this study was to determine (immunohistochemically) estrogen receptor (ER-alpha) expression in vaginal tissue of healthy bitches and in the vaginal and neoplastic tissues of TVT-affected bitches. Fifty-eight bitches were divided into two groups: tumor group (TVT) and control group (healthy). Canine estrous cycle stages were determined by means of exfoliative vaginal cytology, hormone assays, and macroscopic appearance of ovaries. Samples from vaginal and neoplastic tissues were obtained under general anesthesia, fixed in 10% buffered formaldehyde, embedded in paraffin, and sectioned. Anestrus, proestrus and estrus control females had higher ER-alpha expression than diestrus bitches. Within the tumor group, diestrus bitches had significantly higher ER-alpha expression. Although some samples had expression in the endothelium of blood vessels, no ER-alpha expression was observed in neoplastic tissues. In conclusion, vaginal tissue of tumor and control bitches, under different distinct steroid influences, had different ER-alpha expression, whereas ER-alpha expression was not present in neoplastic tissues.