The COVID-19 pandemic has challenged the entire world, and patients with diabetes mellitus (DM) have been particularly affected. We aimed to evaluate predictors of mortality during the first 30 days of hospitalization in critically ill patients with COVID-19 and comorbid DM. This prospective study included 110 critically ill patients admitted with COVID-19 infection. Thirty-two (29%) patients had a previous diagnosis of DM. Clinical variables, laboratory tests, and vascular biomarkers, such as VCAM-1, syndecan-1, ICAM-1, angiopoietin-1, and angiopoeitin-2, were evaluated after intensive care unit (ICU) admission. A comparison was made between patients with and without DM. No difference in mortality was observed between the groups (48.7 vs 46.9%, P=0.861). In the multivariate Cox regression analysis, VCAM-1 levels at ICU admission (HR: 1 [1-1.001], P<0.006) were associated with death in patients with DM. Among patients with DM, advanced age (HR 1.063 [1.031-1.096], P<0.001), increased Ang-2/Ang-1 ratio (HR: 4.515 [1.803-11.308] P=0.001), and need for dialysis (HR: 3.489 [1.409-8.642], P=0.007) were independent predictors of death. Higher levels of VCAM-1 in patients with DM was better at predicting death of patients with severe COVID-19 and comorbid DM, and their cut-off values were useful for stratifying patients with a worse prognosis. Vascular biomarkers VCAM-1 and Ang-2/Ang-1 ratio were predictors of death in patients with severe COVID-19 and comorbid DM and those without DM. Additionally, kidney injury was associated with an increased risk of death.

Glial cells have been implicated in temporal lobe epilepsy in humans and in its models. Astrocytes are lost in several brain regions after acute seizures induced by pilocarpine and may suffer hyperplasia at subsequent time points. This study investigated the effect of N-methyl-(2S,4R)-trans-4-hydroxy-L-proline (NMP) on astrocytes exposed to cytotoxic concentrations of pilocarpine. Astrocytes were incubated with pilocarpine (half maximal inhibitory concentration (IC50)=31.86 mM) for 24 h. Afterwards, they were treated with NMP at concentrations ranging from 3.12 to 100 μg/mL for 24 h. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cytoplasmic reactive oxygen species (ROS) and mitochondrial transmembrane potential (ΔΨm) were analyzed by flow cytometry using 2',7'-dichlorofluorescein diacetate (DCFH-DA) and rhodamine-123 (Rho123), respectively. Expression of glial fibrillary acidic protein (GFAP) and voltage-dependent anion channel-1 (VDAC-1) were measured by western blot. Pilocarpine significantly decreased cell viability and mitochondrial potential and increased ROS concentration significantly by 6.7 times compared to the control. NMP concentrations ≥25 µg/mL protected astrocytes against pilocarpine-induced injury in a concentration-dependent manner. Concomitantly, NMP reduced cytoplasmic ROS accumulation to 27.3, 24.8, and 12.3% in the groups treated with 25, 50, and 100 µg/mL NMP, respectively. NMP also protected mitochondria from pilocarpine-induced depolarization. These effects were associated with improvement of pilocarpine-induced GFAP and VDAC-1 overexpression, which are important biomarkers of astrocyte dysfunction. In conclusion, the improvement of ROS accumulation, VDAC-1 overexpression, and mitochondrial depolarization are possible mechanisms of the NMP protective action on reactive astrocytes.

Acute coronary syndromes are associated with a high prevalence of complications including heart failure (HF). The aim of this study was to investigate the association of novel biomarkers with the occurrence of post-acute myocardial infarction (AMI) HF. A prospective study was conducted with patients admitted to the emergency department with ST-segment elevation myocardial infarction (STEMI). Blood and urine samples were collected for analysis of traditional and novel biomarkers, including interleukin-6, vascular cell adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1). We compared the levels of these biomarkers between patients with and without post-STEMI HF. A total of 48 patients were assessed, with a prevalence of males. Fifteen patients (31.2%) had post-STEMI HF. Patients with HF had higher mean values of IL-6, VCAM-1, and ICAM-1 compared to those who did not develop HF (57.06 vs 14.03 pg/mL, P=0.001; 1719.58 vs 1304.34 ng/mL, P=0.001; and 1594.20 vs 1158.74 ng/mL, P<0.001, respectively). The three biomarkers were shown to be good predictors of post-STEMI HF (IL-6: AUC 0.786, P=0.002; VCAM-1: AUC 0.797, P=0.001; and ICAM-1: AUC 0.825, P<0.0001), with the respective cutoff points being calculated based on the best sensitivity and specificity indexes (IL-6: 8.67 pg/mL; VCAM-1: 1501.42 ng/mL; and ICAM-1: 1262.38 ng/mL). Of the three biomarkers, only VCAM-1 and ICAM-1 had a direct linear association between them (r=0.470, P<0.0001). IL-6, VCAM-1, and ICAM-1 were associated with the development of new post-AMI HF symptoms, but only VCAM-1 and ICAM-1 correlated with each other, possibly because they have the same pathophysiological mechanism of action.

Excess weight (overweight and obesity) is associated with kidney and cardiovascular disease. The aim of this study was to investigate the association between syndecan-1 and renal function among adolescents with excess weight. A total of 56 students from a public school at Fortaleza, CE, Brazil, were investigated. The adolescents were submitted to anthropometric evaluation, including weight, height, blood pressure and body mass index. Blood and urine samples were collected for the determination of serum lipids (total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, triglycerides), and the endothelial injury biomarker syndecan-1. Participants' mean age was 16±1 years (range 14-19 years), and 68% were females. Overweight was observed in 4 cases (7.1%) and obesity in 7 (12.5%). Changes in serum lipid levels were more frequent in the overweight group. A positive correlation between syndecan-1 and serum creatinine (r=0.5, P=0.001) and triglycerides (r=0.37, P=0.004), and a negative correlation with glomerular filtration rate (r=-0.33, P=0.02) were found. These findings suggest that adolescents with excess weight present incipient changes at the cellular level that make them more vulnerable to the development of kidney and cardiovascular diseases.

O objetivo do presente estudo foi avaliar a contagem de células somáticas, a prevalência e a etiologia da mastite bubalina nas estações seca e chuvosa em um rebanho de bubalinos do município de Analândia, estado de São Paulo, Brasil. Adicionalmente, verificou-se o perfil de sensibilidade antimicrobiana dos micro-organismos isolados nas amostras de leite das búfalas. Foram avaliadas 1.042 amostras de leite de búfalas da raça Murrah pertencentes a uma propriedade rural localizada no município de Analândia-SP, obtidas no período de maio de 2011 e novembro de 2012. Após o exame físico da glândula mamária, foram realizados o teste da caneca de fundo escuro e o California Mastits Test (CMT); em seguida, foram colhidas, de forma asséptica, amostras de 50mL de leite de cada quarto mamário, para a contagem de células somáticas (CCS) em aparelho automático e exame microbiológico. Também, foi avaliado o perfil de sensibilidade antimicrobiana a ampicilina, cefoperazona, ceftiofur, enrofloxacina, gentamicina, neomicina, oxacilina, penicilina e sulfametoxazol/trimetoprim, pelo método de difusão em disco. A temperatura média e o índice pluviométrico mensais foram obtidos no Centro Integrado de Informações Agrometeorológicas (CIIAGRO) do Instituto Agronômico de Campinas (IAC). Notou-se que as amostras de leite com resultado positivo no exame microbiológico apresentaram CCS média de 137.720 células/mL, no período seco, e 190.309 células/mL, no período chuvoso. Embora tenha se constatado maior índice de isolamentos de micro-organismos nas amostras de leite obtidas no período chuvoso (69/600) do que no período seco (50/442), a frequência de isolamentos não foi influenciada significativamente pela estação do ano. Os principais gêneros de micro-organismos isolados durante o período seco foram Staphylococcus coagulase negativa (38,4%), Streptococcus agalactiae (28,8%) e Bacillus spp. (7,56%), e no período chuvoso Corynebacterium sp. (23,5%), Streptococcusspp. (32,3%) e Streptococcus agalactiae (9,24%). Verificou-se resistência a três ou mais antimicrobianos em 30,1% dos micro-organismos isolados.

The aim of this study was to evaluate somatic cell count (SCC), prevalence and etiology of mastitis in a dairy buffalo herd from Analândia, São Paulo State, Brazil, in the dry and rainy seasons. Additionally, antimicrobial susceptibility profile of microorganisms isolated from milk samples was also evaluated. 1,042 milk samples from female Murrah buffaloes in a dairy farm located in Analândia, São Paulo State, Brazil, collected between May 2011 and November 2012 were analyzed. After the mammary gland physical examination, strip cup test and California Mastitis Test (CMT) were performed. Afterwards, 50mL of milk samples from each mammary quarter were collected aseptically for SCC in automatic equipment and microbiological examination. The antimicrobial sensitivity profile to ampicillin, cefoperazone, ceftiofur, enrofloxacin, gentamicin, neomycin, oxacillin, penicillin, and sulfamethoxazole/trimethoprim was evaluated by disk diffusion method. The monthly average temperature and pluviometric index were obtained from "Centro Integrado de Informações Agrometeorológicas" (CIIAGRO) of "Instituto Agronômico de Campinas" (IAC). Milk samples with positive results in the microbiological test showed average SCC of 137,720 cells/mL in the dry period and 190,309 cells/mL in the rainy period. Although a higher number of isolated microorganisms was observed in buffalo milk samples during the rainy period (69/600) compared to the dry period (50/442), the season had no significant effect on the frequency of isolation of microorganisms. The main genera of microorganisms isolated were coagulase-negative Staphylococcus (38.4%), Streptococcus agalactiae(28.8%), and Bacillus spp. (7.56%) during the dry season and Corynebacterium sp. (23.5%), Streptococcus spp. (32.3%), and Streptococcus agalactiae (9.24%) during the rainy period. Multidrug resistance was observed in 30.1% of the isolated microorganisms.

RESUMO O leite integral e a bebida láctea UAT são submetidos ao mesmo tratamento térmico, mas são produtos diferentes, já que na bebida láctea é permitida a adição de até 50% de soro lácteo. Neste estudo, objetivou-se avaliar as características microbiológicas destes produtos com relação à legislação vigente do Ministério da Agricultura, Pecuária e Abastecimento (MAPA) e, ainda, da população psicrotrófica, ao longo do período de validade. Foram analisadas 150 amostras, 75 de cada produto, a partir de cinco diferentes marcas comerciais nacionais. As 15 amostras de cada marca foram subdivididas em três lotes: início, meio e final do período de validade, cada qual com cinco amostras do mesmo lote. Verificou-se para o leite que todas as amostras de leite UAT estavam de acordo com a legislação vigente. A população de psicrotróficos aumentou em 26,6% das amostras depois que foram abertas e mantidas sob refrigeração por 48 horas. Os resultados para a bebida láctea evidenciaram que 12% das amostras estavam em desacordo com o estabelecido pela legislação vigente. A população de psicrotróficos aumentou em 36% das amostras depois que foram abertas e mantidas sob refrigeração por 48 horas. Tais resultados devem servir de alerta para a necessidade de fiscalização mais rigorosa da bebida láctea UAT, pois parte das amostras não atenderam aos padrões microbiológicos estabelecidos pela legislação.

ABSTRACT UHT milk and dairy drinks are submitted to the same heat treatment, but they are different products, since the addition of up to 50% of whey is allowed in dairy drinks. The aim of this study was to evaluate the microbiological characteristics of these products in relation to current legislation, during the course of their shelf life. In the present study, 150 samples, 75 of each type of product, from 5 different national brands were analyzed. The 15 samples of each brand were separated into 3 lots containing 5 samples each, analyzed at the beginning, middle and end of the validity period. Upon microbiological analysis all UHT milk samples were in conformance with current legislation. After the samples were opened and chilled for 48 hours, 26.6% of them presented an increased psychrotrophic population. The dairy drink results showed that 12% of the samples were out of conformance with the current legislation. After the samples were opened and chilled for 48 hours, 36% of them presented an increased psychrotrophic population. These results should be considered as an alert in regard to the need for rigorous inspection of UHT milk and dairy drink products.

Pesquisaram-se a presença de Bacillus cereus e a produção de enterotoxinas produzidas por esses microrganismos em 120 amostras de diversos tipos de leite. Bacillus cereus foi isolado e identificado em 22 (73,3%), 15 (50,0%), 29 (96,7%) e quatro (13,3%) amostras de leite em pó, cru, pasteurizado e UAT (longa vida), respectivamente. Para a detecção de enterotoxinas pela técnica da alça ligada de coelho, foram positivos, respectivamente, três (13,6%), um (7,1%) e 10 (35,7%) microrganismos isolados das amostras de leite em pó, leite cru e leite pasteurizado. Pelo teste de aumento de permeabilidade vascular, dois (9,1%), um (7,1%), um (3,6%) e um (4,0%) microrganismos isolados de leite em pó, cru, pasteurizado e UAT apresentaram-se enterotoxigênicos, respectivamente. O uso da técnica de aglutinação passiva em látex demonstrou a produção da toxina diarréica por três (33,3%), sete (63,6%), quatro (30,8%) e oito (80,0%) microrganismos isolados, respectivamente, de leite em pó, cru, pasteurizado e UAT. Os resultados indicam um risco potencial, podendo colocar em risco a saúde dos consumidores desses produtos.

A hundred and twenty samples of different types of milk were examined to the presence of Bacillus cereus and the enterotoxigenic potential of the isolated strains. Bacillus cereus was isolated and identified in 22 (73.0%), 15 (50.0%), 29 (96.7%) and four (13.3%) samples of powder, raw, pasteurized and UHT milk, respectively. The enterotoxigenicity detection using the rabbit ileal loop assay showed positive, respectively, three (13.6%), one (7.1%) and 10 (35.7%) isolated strains from powder, raw and pasteurized milk. Using vascular permeability activity assay two (9.1%), one (7.1%), one (3.6%) and one (4.0%) isolated strains from powder, raw, pasteurized and UHT milk were positive, respectively. The reversed passive latex agglutination test showed diarrheal toxin production by three (33.3%), seven (63.6%), four (30.8%) and eight (80.0%) strains isolated from, respectively, powder, raw, pasteurized and UHT milk. Those results indicated a potential hazard and may put in risk the health of the consumers of these products.

RESUMO O objetivo do trabalho foi avaliar a resistência antimicrobiana de cepas de Salmonella isoladas de águas de escaldamento, evisceração e resfriamento; de carcaças não evisceradas, evisceradas e resfriadas; penas e fezes frente à ação de antimicrobianos de uso comum. Foram coletadas 288 amostras e isoladas 29 cepas de Salmonella spp., dentre elas S. Kentucky (34,5%); S. Enteritidis (20,8%); S. Anatum e S. Enterica subsp. Enterica 8,20:-:z6 (13,8%); S. Typhimurium (6,9%);S. Enterica subsp. Enterica 4,5, 12:i:-, S. Saintpaul e S. Tennessee 3,4%. O teste de sensibilidade frente a 12 agentes antimicrobianos foi realizado com as amostras de Salmonella spp. isoladas e seguiu a metodologia de Kirby-Bauer, 25 (86,2%) amostras foram resistentes ao aztreonam e à ampicilina, 21 (72,4%) à tetraciclina e 16 (55,2%) à amoxicilina/ácido clavulânico e sulfazotrim. Os isolados apresentaram menor resistência à gentamicina , com uma amostra (3,45%) resistente, duas (6,9%) amostras foram resistentes à amicacina. Nenhuma das amostras testadas apresentou 100% de resistência ou sensibilidade aos antimicrobianos utilizados. As amostras de Salmonella spp. foram em grande proporção resistentes aos princípios antimicrobianos normalmente utilizados em avicultura, estes dados servem como alerta contra o uso indiscriminado de antibióticos, que pode contribuir para a seleção de cepas resistentes, agentes de doenças transmissíveis por alimentos em seres humanos.

ABSTRACT The aim of this study was to evaluate the level of antibiotic resistance of Salmonella strains isolated from scald, evisceration, and chiller water; non-eviscerated, eviscerated, and chilled carcasses; feathers; and feces collected in six chicken abattoirs. The 29 Salmonella strains belonged to S. Kentucky (34.5%); S. Enteritidis (20.8%); S. Anatum e S. Enterica subsp. Enterica 8,20:-:z6 (13.8%); S. Typhimurium (6.9%); S. Enterica subsp. Enterica 4,5, 12:i:-, S. Saintpaul and S. Tennessee (3.4%). All strains were submitted to antibiotic resistance test following Kirby-Bauer methodology, 25 (86.2%) strains were resistant to aztreonam and to ampicilin, 21 (72.4%) to tetracycline and 16 (55.2%) to amoxicilin/clavulanic acid and to sulfazotrim. Only one strain (3.45%) was gentamicin resistant. Two strains (6.9%) were resistant to amicacin. Resistance and susceptibility to all antibiotics tested was not detected.Salmonella strains were total or partially resistant to the antibiotics frequently used in aviculture. These note-worthy data draw attention to a danger of indiscriminate use of antibiotics in aviculture that may contribute for selection of resistant strains and this can cause foodborne infections in humans.

RESUMO No período de julho a dezembro de 2002, foi avaliada a qualidade microbiológica de 60 amostras de queijo minas frescal, através do Número Mais Provável (NMP) de coliformes fecais, contagem de Staphylococcus coagulase positiva, pesquisa de Salmonella spp., pesquisa de Listeria monocytogenes e pesquisa de Campylobacter spp., sendo que, 30 amostras eram de produção artesanal e 30 de produção industrial fiscalizadas pelo Serviço de Inspeção Estadual e Federal e foram adquiridas no comércio do Município de Jaboticabal, São Paulo. Quanto a presença de coliformes fecais, apresentaram-se em desacordo com o estabelecido pela Agência Nacional de Vigilância Sanitária (ANVISA) 83,4% das amostras artesanais e para as amostras industriais 66,7%. Para os valores obtidos na contagem de Staphylococcus coagulase positiva, 20% das amostras artesanais e 10% das amostras industriais, apresentaram-se em desacordo com o estabelecido pela Anvisa. Com relação a detecção de Salmonella spp., Listeria monocytogenes e Campylobacter spp., tais microrganismos não foram isolados em 25 g do produto, mostrando que em relação a estes agentes os queijos analisados tanto de produção artesanal como os industrializados, apresentaram-se dentro dos padrões estabelecidos (ausência em 25 g do produto). Os resultados microbiológicos revelaram que 86,7% das amostras artesanais e 66,7% das amostras industriais encontravam-se em desacordo com os padrões estabelecidos pela Anvisa, segundo a Resolução n° 12 de 02 de janeiro de 2001. Tais resultados demonstram o risco potencial que este produto pode representar para a saúde da população consumidora.

ABSTRACT From July to December 2002 the microbial quality of 60 samples of “minas frescal” cheese was analyzed, from which 30 hand made samples and 30 from dairy industry inspected by Federal Inspection Service. The microbiological results revealed that 86.7% from hand-made samples and 66.7% from dairy industry samples were not in standards permitted by the current legislation (Agência Nacional de Vigilância Sanitária – ANVISA), according to resolution number 12 of January 2nd, 2001. Of the hand made and dairy industry samples, 83.4% and 66.7% (> 5.0 x 102 MPN/g), respectively, were out of the standards for contamination by fecal colliforms. The values obtained by coagulase positive staphylococci count, 20% of the hand made samples and 10% of the dairy industry samples were not accordind to the standards permitted (5.0 x 102 CFU/g). As for Salmonella spp., Listeria monocytogenes and Campylobacter spp., they were not detected in 25 g of the product, showing that, concerning these organisms, both sources of samples were according to ANVISA standards (absence in 25 g of the product). Our results reveal the potential risk to public health of the marketable “minas frescal” cheese when sanitation care is deficient.

RESUMO Objetivando estudar as características microbiológicas do leite UAT (ultra alta temperatura) ao longo de sua industrialização, foram analisadas 60 amostras de leite cru, 60 de pasteurizado e 30 de UAT, colhidas em uma linha de produção. Verificou-se que as populações médias de microrganismos mesófilos, psicrotróficos e esporos diminuiram com o tratamento térmico, pois as contagens, respectivamente, da ordem de 108, 107 e 104 UFC/mL no leite cru, caíram para 104, 102 e 102 UFC/mL no pasteurizado e para inferior a 1,0, 10,0 e 1,0 UFC/mL no leite UAT. O mesmo foi verificado com a população de microrganismos termófilos, que no leite pasteurizado era da ordem de 103 UFC/mL e no UAT inferior a 1,0 UFC/mL. Os resultados permitem concluir que o processamento térmico aplicado à matéria-prima para a produção do leite UAT foi capaz de reduzir, mas não de eliminar a carga microbiana inicial.

ABSTRACT With the aim of studying microbiological characteristics of UHT milk (ultra high temperature) during the industrialization, the present study analysed 60 samples of raw milk, 60 of pasteurized and 30 of UHT process, collected ata a factory. It was seen that the population of mesophilic, psychrotrophic and sporulated microrganisms decreased with thermal threatment, because the number of microrganisms were 108, 107 and 104 CFU/mL in raw milk, and decreased to 104, 102 and 105 CFU/mL in pasteurized to less tham 1.0, 10.0 and 1.0 CFU/mL in UHT milk. The same was verified with the mesophilic population of thermophils, which, in the pasteurized milk were 103 CFU/mL and UHT 1.0 CFU/mL. The results allow for the conclusion that termal threatment applied to raw material in the production of UHT milk were able to reduce but not to eliminate the initial number of microrganisms.

RESUMO Os produtos de origem avícola têm sido implicados como veículos de bactérias patogênicas. Desta maneira, a qualidade microbiológica desses produtos deve apresentar condições apropriadas para o consumo. Com a finalidade de verificar a qualidade higiênico-sanitária dos produtos avícolas disponíveis no comércio, foram analisadas 47 amostras de produtos avícolas, sendo 5 amostras de coxa/sobrecoxa, 5 conjuntos de fígado e moela, 15 amostras de CMS, 7 de lingüiça, 5 de salsicha, 5 de hambúrguer e 5 amostras de peito empanado, as quais foram submetidas a contagem padrão em placas de microrganismos heterotróficos aeróbios mesófilos e psicrotróficos, bem como a determinação do número mais provável de coliformes totais e coliformes fecais. Os resultados das amostras foram apresentados por meio dos valores mínimo e máximo observados para cada grupo de microrganismos estudado em suas amostras correspondentes. Levando-se em consideração que a legislação brasileira vigente determina limite máximo apenas para a presença de coliformes fecais, a presente pesquisa detectou uma amostra de miúdos (fígado e moela) (2,1%) e 3 amostras de lingüiça (6,4%) em desacordo com os padrões estabelecidos para esse grupo de microrganismos, demonstrando que, das 47 amostras analisadas, apenas quatro amostras (8,5%) apresentaram-se em desacordo com o padrão estabelecido.

ABSTRACT Products from poultry sources can be a route of introduction of pathogenic bacterium. Therefore the microbiological quality of these products should be appropriate for consumption. With the aim to establish the sanitary-hygiene quality of the poultry products available at the commercial facilities 47 samples of the following poultry products were analyzed: 5 thigh samples, 5 samples of liver together with gizzard, 15 mechanically boned meat samples, 7 sausage samples, 5 hotdog sausage samples, 5 hamburger samples, and 5 covered chest samples. These samples were submitted to counting standard in plates of mesophilic and psychrophilic aerobic heterotrophic microorganisms, as well as determination of the most probable number of total and fecal coliforms.The results of the analized samples were shown in minimum and maximum values observed for each microorganism and for each chicken product. According to Standard Brazil Procedures only the maximum value of fecal coliforms is determined; this study detected one sample (2.1%) of liver and gizzard and 3 sausage samples (6.4%) out of the standard established by the legislation for these microorganisms, showing that from the 47 analyzed samples only four (8.5%) were out of the standard.

Cento e dez amostras de 11 diferentes marcas de leite ultra alta temperatura (UAT), comercializadas em São José do Rio Preto - SP, foram submetidas à contagem de microrganismos heterotróficos mesófilos viáveis e à pesquisa de bactérias do grupo do Bacillus cereus. A população de microrganismos mesófilos variou de <1,0<FONT FACE=Symbol>´</FONT>10² UFC/ml a >1,0<FONT FACE=Symbol>´</FONT>10(6) UFC/ml. Bactérias do grupo do Bacillus cereus foram verificadas em 13 (11,8%) amostras. Os resultados evidenciaram elevada população de microrganismos indicadores mesófilos.

One hundred and ten ultra high temperature (UHT) milk samples, from 11 different brands retailed in São José do Rio Preto - SP were analysed for mesophilic heterotrophic microorganisms counting, as well as for the presence of Bacillus cereus. Mesophilic heterotrophic microorganisms ranged from <1.0<FONT FACE=Symbol>´</FONT>10² CFU/ml to >1.0<FONT FACE=Symbol>´</FONT>10(6) CFU/ml. Bacillus cereus was detected in 13 (11.8%) samples. The results demonstrated high countings of mesophilic microorganisms.

Because thalidomide and pentoxifylline inhibit the synthesis and release of tumor necrosis factor-alpha (TNF-alpha), we determined the effect of these drugs on the renal damage induced by supernatants of macrophages activated with Crotalus durissus cascavella venom in order to identify the role of TNF-alpha in the process. Rat peritoneal macrophages were collected with RPMI medium and stimulated in vitro with C.d. cascavella venom (10 µg/ml) in the absence and presence of thalidomide (15 µM) or pentoxifylline (500 µM) for 1 h and washed and kept in culture for 2 h. Supernatant (1 ml) was tested on an isolated perfused rat kidney (N = 6 for each group). The first 30 min of each experiment were used as control. The supernatant was added to the perfusion system. All experiments lasted 120 min. The toxic effect of the preparation of venom-stimulated macrophages on renal parameters was determined. At 120 min, thalidomide (Thalid) and pentoxifylline (Ptx) inhibited (P < 0.05) the increase in perfusion pressure caused by the venom (control = 114.0 ± 1.3; venom = 137.1 ± 1.5; Thalid = 121.0 ± 2.5; Ptx = 121.4 ± 4.0 mmHg), renal vascular resistance (control = 4.5 ± 0.2; venom = 7.3 ± 0.6; Thalid = 4.5 ± 0.9; Ptx = 4.8 ± 0.6 mmHg/ml g-1 min-1), urinary flow (control = 0.23 ± 0.001; venom = 0.44 ± 0.01; Thalid = 0.22 ± 0.007; Ptx = 0.21 ± 0.009 ml g-1 min-1), glomerular filtration rate (control = 0.72 ± 0.06; venom = 1.91 ± 0.11; Thalid = 0.75 ± 0.04; Ptx = 0.77 ± 0.05 ml g-1 min-1) and the decrease in percent tubular sodium transport (control = 77.0 ± 0.9; venom = 73.9 ± 0.66; Thalid = 76.6 ± 1.1; Ptx = 81.8 ± 2.0%), percent tubular chloride transport (control = 77.1 ± 1.2; venom = 71.4 ± 1.1; Thalid = 77.6 ± 1.7; Ptx = 76.8 ± 1.2%), and percent tubular potassium transport (control = 72.7 ± 1.1; venom = 63.0 ± 1.1; Thalid = 72.6 ± 1.0; Ptx = 74.8 ± 1.0%), 30 min before and during the stimulation of macrophages with C.d. cascavella venom. These data suggest the participation of TNF-alpha in the renal effects induced by supernatant of macrophages activated with C.d. cascavella venom.

Many studies have reported the occurrence of lethal acute renal failure after snakebites. The aim of the present investigation was to determine alterations in renal function produced by Crotalus durissus terrificus venom and crotoxin as well as the histological alterations induced by these venoms. Isolated kidneys from Wistar rats weighing 240 to 280 g were perfused with Krebs-Henseleit solution containing 6 g% of previously dialyzed bovine serum albumin. The effects of Crotalus durissus terrificus venom and crotoxin were studied on glomerular filtration rate (GFR), urinary flow (UF), perfusion pressure (PP) and percentage sodium tubular transport (%TNa+). The infusion of Crotalus durissus terrificus venom (10 µg/ml) and crotoxin (10 µg/ml) increased GFR (control80 = 0.78 ± 0.07, venom80 = 1.1 ± 0.07, crotoxin80 = 2.0 ± 0.05 ml g-1 min-1, P<0.05) and UF (control80 = 0.20 ± 0.02, venom80 = 0.32 ± 0.03, crotoxin80 = 0.70 ± 0.05 ml g-1 min-1, P<0.05), and decreased %TNa+ (control100 = 75.0 ± 2.3, venom100 = 62.9 ± 1.0, crotoxin80 = 69.0 ± 1.0 ml g-1 min-1, P<0.05). The infusion of crude venom tended to reduce PP, although the effect was not significant, whereas with crotoxin PP remained stable during the 100 min of perfusion. The kidneys perfused with crude venom and crotoxin showed abundant protein material in the urinary space and tubules. We conclude that Crotalus durissus terrificus venom and crotoxin, its major component, cause acute nephrotoxicity in the isolated rat kidney. The current experiments demonstrate a direct effect of venom and crotoxin on the perfused isolated kidney.