Predictors of mortality in critically ill patients with COVID-19 and diabetes.

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.

Predictors of mortality in critically ill patients with COVID-19 and diabetes

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.

N-Methyl-(2S, 4R)-trans-4-hydroxy-L-proline, the major bioactive compound from Sideroxylon obtusifolium, attenuates pilocarpine-induced injury in cultured astrocytes.

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.

Kidney injury molecule-1 and urinary gamma-glutamyl transferase as biomarkers of acute kidney injury in cats.

OBJECTIVES: To evaluate the concentration of kidney injury molecule-1 and activity of urinary gamma-glutamyl transferase in cats with urethral obstruction and healthy cats. MATERIALS AND METHODS: Blood and urine samples were collected from a group of 15 healthy cats (control group) and a group of 20 cats with urethral obstruction at presentation, and 24 hours and 7 days after unblocking the obstruction. The serum creatinine, urinary creatinine and urinary gamma-glutamyl transferase were measured by spectrophotometry and kidney injury molecule-1 by the sandwich enzyme-linked immunosorbent assay. RESULTS: On presentation, cats with obstruction had serum creatinine concentration and urinary gamma-glutamyl transferase index higher than healthy cats (mean difference 544 µmol/L, 95% confidence intervals 222 to 865 µmol/L, and 0.0022 U/µmol-uCre, 0.00043 to 0.0039 U/µmol-uCre, respectively), urine creatinine concentration lower (mean difference 25,624 µmol/L, 17,329 to 33,919 µmol/L), and no significant difference in the kidney injury molecule-1/urinary creatinine ratio (mean difference 13 pg/µmol-uCre, -33 to 59 pg/µmol-uCre). In the group of cats with urinary obstruction, over time serum creatinine decreased, urine creatinine increased, urinary gamma-glutamyl transferase index did not change significantly, and kidney injury molecule-1/urinary creatinine ratio increased. CLINICAL SIGNIFICANCE: Cats with post-renal obstruction and potential intrinsic renal damage had higher urinary gamma-glutamyl transferase index than healthy cats at the time of presentation and showed increase in kidney injury molecule-1/urinary creatinine ratio over time.

N-Methyl-(2S, 4R)-trans-4-hydroxy-L-proline, the major bioactive compound from Sideroxylon obtusifolium, attenuates pilocarpine-induced injury in cultured astrocytes

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.

Interleukin-6 and adhesion molecules VCAM-1 and ICAM-1 as biomarkers of post-acute myocardial infarction heart failure.

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.

Renal Alterations Induced by the Venom of Colombian Scorpion Centruroides Margaritatus.

BACKGROUND: Scorpion venom causes renal injury and affects vascular ion-channels function. Centruroides margaritatus scorpion is found in Colombia and is frequently the cause of envenomation accidents; however, its renal impact has never been investigated. OBJECTIVE: To evaluate the effects of C. margaritatus venom (CmV) on renal parameters using isolated rat kidney and renal cell culture models. METHODS: Wistar rats (n = 5, weighing 240-300 g) were first perfused with Krebs-Henseleit solution containing 6 g 100 mL-1 bovine serum albumin. After 30 minutes, the kidneys were perfused with CmV to a final concentration of 10 µgmL-1; evaluation was performed by measuring Perfusion Pressure (PP), Renal Vascular Resistance (RVR), Urinary Flow (UF), Glomerular Filtration Rate (GFR), and percentage of electrolyte tubular transport. Moreover, kidney histological analyses and cell cytotoxicity in renal tubule epithelial cells (MDCK) and proximal tubular cells (LLC-MK2) were assessed. RESULTS: CmV increased PP and RVR 60 min after perfusion. On the other hand, UF, GFR, and the percentages of sodium, potassium and chloride tubular transport decreased after experimental envenomation. UF dropped after 120 min, while GFR and percentage of electrolyte tubular transport diminished after 60, 90 and 120 min. CmV was not toxic to MDCK cell line but reduced the viability of LLC-MK2 cells at concentrations ranging from 6.25 to 200 µgmL-1. Histological analyses disclosed hydropic degeneration, edema, and protein deposits. Flow cytometry disclosed that cell death occurred predominantly by necrosis. CONCLUSION: Our results suggest that C. margaritatus venom can trigger renal impairment, mainly in the proximal kidney tubule.

Antichagasic effect of violacein from Chromobacterium violaceum.

AIMS: Violacein (VIO), a bacterial pigment produced by Chromobacterium violaceum, was examined to evaluate the antichagasic activity and its action mechanism against Trypanosoma cruzi Y strain. METHODS AND RESULTS: Violacein was tested against the epimastigote, trypomastigote and amastigote forms of T. cruzi Y strain (benznidazole-resistant strain). VIO inhibited all T. cruzi developmental forms, including amastigotes, which is implicated in the burden of infection in the chronic phase of Chagas disease (CD). VIO induced cell death in T. cruzi through apoptosis, as determined by flow cytometry analyses with specific molecular probes and morphological alterations, such as involvement of reactive oxygen species and changes in mitochondrial membrane potential and cell shrinkage. CONCLUSION: The results suggest antichagasic activity of VIO against T. cruzi Y strain with apoptotic involvement. SIGNIFICANCE AND IMPACT OF THE STUDY: The treatment of CD has limited efficacy and side effects that restrict patient tolerability and compliance. The VIO molecule could be used as a model for therapeutic alternatives for this disease.

Interleukin-6 and adhesion molecules VCAM-1 and ICAM-1 as biomarkers of post-acute myocardial infarction heart failure

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.

Association between syndecan-1 and renal function in adolescents with excess weight: evidence of subclinical kidney disease and endothelial dysfunction.

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.

Influence of ßS-Globin Haplotypes and Hydroxyurea on Arginase I Levels in Sickle Cell Disease.

Introduction. Sickle cell disease (SCD) is characterized by hemoglobin S homozygosity, leading to hemolysis and vasoocclusion. The hemolysis releases arginase I, an enzyme that decreases the bioavailability of nitric oxide, worsening the symptoms. The different SCD haplotypes are related to clinical symptoms and varied hemoglobin F (HbF) concentration. The aim of this study was to evaluate the impact of the ßS gene haplotypes and HbF concentration on arginase I levels in SCD patients. Methods. Fifty SCD adult patients were enrolled in the study and 20 blood donors composed the control group. Arginase I was measured by ELISA. The ßS haplotypes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Statistical analyses were performed with GraphPad Prism program and the significance level was p < 0.05. Results. Significant increase was observed in the arginase I levels in SCD patients compared to the control group (p < 0.0001). The comparison between the levels of arginase I in three haplotypes groups showed a difference between the Bantu/Bantu × Bantu/Benin groups; Bantu/Bantu × Benin/Benin, independent of HU dosage. An inverse correlation with the arginase I levels and HbF concentration was observed. Conclusion. The results support the hypothesis that arginase I is associated with HbF concentration, also measured indirectly by the association with haplotypes.

Prevalence and etiology of buffalo mastitis and milk somatic cell count in dry and rainy seasons in a buffalo herd from Analândia, São Paulo State, Brazil / Prevalência e etiologia da mastite bubalina e contagem de células somáticas no leite, nas estações seca e chuvosa, em um rebanho bubalino do município de Analândia, Estado de São Paulo, Brasil

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...

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...

Prevalence and etiology of buffalo mastitis and milk somatic cell count in dry and rainy seasons in a buffalo herd from Analândia, São Paulo State, Brazil / Prevalência e etiologia da mastite bubalina e contagem de células somáticas no leite, nas estações seca e chuvosa, em um rebanho bubalino do município de Analândia, Estado de São Paulo, Brasil

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.(AU)

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%), Streptococcus spp. (32,3%) e Streptococcus agalactiae (9,24%). Verificou-se resistência a três ou mais antimicrobianos em 30,1% dos micro-organismos isolados.(AU)

Antimicrobial effect of Dinoponera quadriceps (Hymenoptera: Formicidae) venom against Staphylococcus aureus strains.

AIMS: Dinoponera quadriceps venom (DqV) was examined to evaluate the antibacterial activity and its bactericidal action mechanism against Staphylococcus aureus. METHODS AND RESULTS: DqV was tested against a standard strain of methicillin-sensitive Staphylococcus aureus (MSSA), Staph. aureus ATCC 6538P and two standard strains of methicillin-resistant Staphylococcus aureus (MRSA), Staph. aureus ATCC 33591 and Staph. aureus CCBH 5330. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), the rate of kill and pH sensitivity of the DqV were determined by microdilution tests. Bactericidal and inhibitory concentrations of DqV were tested to check its action on Staph. aureus membrane permeability and cell morphology. The MIC and MBC of DqV were 6·25 and 12·5 µg ml(-1) for Staph. aureus ATCC 6538P, 12·5 and 50 µg ml(-1) for Staph. aureus CCBH 5330 and 100 and 100 µg ml(-1) for Staph. aureus ATCC 33591, respectively. Complete bacterial growth inhibition was observed after 4 h of incubation with the MBC of DqV. A lowest MIC was observed in alkaline pH. Alteration in membrane permeability was observed through the increase in crystal violet uptake, genetic material release and morphology in atomic force microscopy. CONCLUSIONS: The results suggest antibacterial activity of DqV against Staph. aureus and that the venom acts in the cell membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: Alteration in membrane permeability may be associated with the antimicrobial activity of hymenopteran venoms.

Diagnosis to detect porcine transmissible gastroenteritis virus (TGEV) by optical and transmission electron microscopy techniques / Diagnóstico para detectar el virus de la gastroenteritis porcina transmisible por técnicas de microscopía óptica y electrónica de transmisión

The porcine transmissible gastroenteritis is a highly severe contagious disease, caused by virus of the Coronaviridae family, genus Coronavirus. Its epizootic shape can reach a rate of up to 100 percent mortality in piglets under two weeks of age as a result of severe dehydration. In this study fragments of small intestine and stool samples were collected from 75 autopsied pigs from properties. The samples of the fragments were frozen and sent to the Laboratory of Electron Microscopy, Instituto Biológico, SP, Brazil, for histological and transmission electron microscopic analyses. According to histological H&E technique, atrophy, villous necrosis and destruction of the enterocytes were observed in 35 (46.6 percent) out of the 75 fragments of the small intestine samples. On the immunohistochemistry technique 19 (25.3 percent) fragments were positively stained with DAB in the Ag-Ac reaction (MabTGEV). In 19 (25.3 percent) positive samples analyzed by in situ hybridization, a brown stain of enterocytes was observed, mainly in the epithelial cells of the villi. By the negative staining technique, we visualized enveloped, pleomorphic coronavirus particles, with typical radial projections resembling solar corona, with 140 nm diameter in 21 samples (28 percent) of the small intestine fragments and in 16 (21.3 percent) stool samples. In the ultrathin sections of 21 (28 percent) samples of small intestine, complete coronavirus particles with 80 nm diameter were seen among the microvilli and in the cytoplasm of epithelial cells. Immature particles with 60 nm diameter, budding from cell membrane and from a rough endoplasmic reticulum and also inside the vacuoles were visualized. In 19 (25.3 percent)...

La gastroenteritis transmisible porcina se caracteriza por ser una enfermedad altamente contagiosa y aguda, causada por virus de la familia Coronaviridae, género Coronavirus. Su forma epizoótica puede alcanzar una tasa de hasta 100 percent de mortalidad en lechones con menos de dos semanas de edad, como resultado de deshidratación severa. En este trabajo se recogieron 75 fragmentos de intestino delgado y muestras de heces de 75 cerdos autopsiados. Las muestras se congelaron y se enviaron al Laboratorio de Microscopía Electrónica, del Instituto Biológico, SP, Brasil, para el análisis histológico y por microscopía electrónica de transmisión. Por la técnica histológica de H&E fue observado atrofia y necrosis de la vellosidades además de la destrucción de los enterocitos en 35 (46,6 ppor ciento) de las 75 muestras de fragmentos del intestino delgado. Por inmunohistoquímica 19 (25,3 por ciento) de los fragmentos se tiñeron positivamente por el DAB en la reacción Ag-Ac (MabTGEV). En 19 (25,3 por ciento) muestras positivas mediante hibridación in situ, se observó tinción marrón de los enterocitos, principalmente en las células epiteliales de las vellosidades. Por la técnica de coloración negativa, se observaron partículas de coronavirus, encapsuladas, pleomórficas con proyecciones radiales típicas, en forma de corona solar, midiendo alrededor de 140 nm de diámetro en 21 (28 por ciento) muestras de fragmentos del intestino delgado y en 16 (21,3 por ciento muestras de heces. En cortes ultra finos de 21 (28 por ciento) fragmentos de intestino delgado fueron visualizadas partículas de coronavirus completas con 80 nm de diámetro entre las microvellosidades y en el citoplasma de las células epiteliales y partículas inmaduras de 60 nm de diámetro brotando de las membranas celulares y del retículo endoplasmático rugoso y en el interior de las vacuolas. En 19 (25,3 por ciento)...

Ultrastructural study of bovine papillomavirus during outbreaks in Brazil / Estudio ultraestructural del virus papiloma bovino durante brotes en Brasil

Papillomaviruses, belonging to the Papillomaviridae family, are small oncogenic viruses, causing papillomas and fibropapillomas in the mucosal and cutaneous epithelia of several animals. In bovine species, thirteen types (BPV 1-13) were characterized to date. In this study, the occurrence of papillomatosis in four outbreaks in cattle herds, coming from Brazilian states were registered. The papillomatous lesions were found located in the teats, udders, head and neck. Under the transmission electron microscope, by the negative staining technique, it was possible to visualized rounded-format papillomavirus, with icosahedral symmetry, characterized as "full" and "empty" particles, measuring on average 60 nm in diameter, in all the 40 samples observed of skin lesion fragments. The ultrathin sections revealed the presence of groups of viral, intranuclear, rounded particles measuring 35 nm in diameter and tubular particles with a diameter of 35-39 nm. At immunoelectron microscopy technique, positivity obtained was marked by the presence of aggregates of viral particles formed by the antigen-antibody interaction. In the immunocytochemistry technique, the antigen-antibody reaction showed colloidal gold particles evenly distributed over the surface of the virus. These results showed the importance of the transmission electron microscopy techniques in the diagnosis of bovine papillomatosis that can be used in routine procedures to identify viral agent of this important disease.

Los virus del papiloma que pertenecen a la familia Papillomaviridae son pequeños virus oncogénicos que causan papilomas y fibropapilomas en epitelio cutáneo y mucoso de distintas especies de animales. En el ganado vacuno, trece tipos (BPV- 1-13) se caracterizaron hasta el momento. En este estudio, se documenta la ocurrencia de cuatro brotes de papilomatosis en los rebaños de ganado, procedentes de estados brasileños. Las lesiones papilomatosas se localizaron en los pezones, la ubre, la cabeza y el cuello. Al microscopio electrónico de transmisión, en la técnica de tinción negativa fueran visualizadas partículas del virus del papiloma redondeadas, con simetría icosaédrica, caracterizadas como "llenas" y "vacías", midiendo unos 60 nm de diámetro en todas las 40 muestras de fragmentos de lesión de piel estudiado. Los cortes ultra finos mostraron la presencia de grupos de partículas virales, intranucleares redondeadas con 35 nm de diámetro y tubulares 35-39 nm de diámetro. En la técnica de microscopía inmunoelectrónica, la positividad obtenida se caracterizó por la presencia de agregados de partículas virales formadas por la interacción antígeno-anticuerpo. En la aplicación de la técnica de inmunocitoquímica, la reacción antígeno-anticuerpo mostró partículas de oro coloidal distribuidos de manera uniforme sobre la superficie del virus. Estos resultados muestran la importancia de las técnicas de microscopía electrónica de transmisión en el diagnóstico de papilomatosis bovina, que pueden ser utilizados en los procedimientos de rutina para la identificación del agente viral causante de esta importante enfermedad.

Diminuição da contagem de células somáticas em leite cru através de modelo experimental de fraude / Decrease of somatic cell count in raw milk through experimental model of fraud

Tendo em vista busca por rendimento industrial e lucratividade, as indústrias propuseram pagamento pela qualidade do leite produzido, assim o produtor receberia bonificação ou penalização referentes à composição do leite como teores de gordura, proteína e contagem de células somáticas (CCS). Porém, quando existem programas de melhoria de qualidade onde o produtor poderá ser bonificado/penalizado ocorre o risco de fraudes surgirem, a fim de atender o estabelecido. O objetivo desse estudo foi verificar a viabilidade do modelo experimental de fraude mediante a diminuição da CCS pela subtração de porções de sobrenadante de leite mantido em estação e avaliar as consequências desta subtração sobre alguns componentes do leite. Para tal, foram utilizadas amostras de leite de 18 animais, submetidas a dois diferentes períodos de estação e/ou repouso (30 e 60 minutos) e posteriormente subtraídos 5,0%, 7,5% e 10,0% dos sobrenadantes do leite oriundos destas vacas. Os resultados demonstraram que é possível diminuir a CCS do leite por meio do modelo experimental de fraude. Através das análises estatísticas pode-se concluir que o melhor modelo de diminuição de CCS é obtido pela subtração de 7,5% do sobrenadante por 30 minutos de repouso. Os programas de pagamento por qualidade devem ser usados como forma de incentivo à melhoria da qualidade do leite e para evitar as fraudes. O pagamento pelo requisito da gordura deve ser mantido para fins de avaliação da amostragem correta do leite e como um indicativo de possíveis fraudes na CCS do leite. Apesar do modelo experimental ter demonstrado a possibilidade da diminuição da CCS, trazendo bonificação ao produtor quanto a este componente, as análises estatísticas demonstraram que o mesmo não é viável, pois os produtores perderiam em outros componentes que também são utilizados como parâmetros para bonificação e em volume de leite, o que poderia gerar a penalização.

In order to reach the industrial profitability the industries proposed the Premium Milk Payments, where the producer would get bonus or would be punished focusing on milks composition like fat, protein and (SCC) somatic cell count. However, when there is improvement milk quality program where the producer can have bonus or being penalized there is a risk of fraud. This study aims to verify the feasibility of the experimental model to SCC fraud by subtraction portions of the supernatant of milk and evaluate the consequences of this subtraction on some components of the milk. For experimental model were used milk samples from 18 lactating cows, and milk samples subjected to two different rest period and subsequently subtracted 5,0%, 7,5% and 10,0% of supernatant of milk. The results showed that is possible to reduce SCC from the milk through the experimental model of fraud described. The statistical analyzes showed as results the best model to reduce SCC obtained by subtracting 7,5% of the supernatant by 30 minutes rest. The Premium Milk Payments should be used as an incentive to improve the quality of milk to prevent fraud. The payment requirement for fat should be kept for the purpose of assessing the correct sampling of milk and as indication of possible fraud in the SCC from milk samples. Despite the experimental model have demonstrated the possibility of decreased SCC, bringing bonus the producer on this component, the statistical analyzes show that it is not feasible, because the produces would lose in other components that are also used as a reference for subsidy and milk volume, which could be lead to penalty.

ISOLAMENTO DE Yersinia enterocolitica EM SUÍNOS AO ABATE / ISOLATION OF Yersinia enterocolitica IN SLAUGHTERED SWINE

O gênero bacteriano Yersinia compreende 14 espécies (BOTTONE et al., 2005), entre as quais, quatro são consideradas patogênicas aos animais e/ou aos humanos, entre elas, a Yersinia enterocolitica (ORTIZ MARTÍNEZ, 2010). Apesar de sua distribuição heterogênea, (LYNCH et al., 2006; BONARDI et al., 2010), a maior fonte de infecção de Y. enterocolitica  para os humanos é o suíno e a principal via de transmissão, a carne suína e seus produtos contaminados (FREDRIKSSON-AHOMAA et al., 2011; DRUMMOND et al., 2012). Segundo Howard et al. (2006) várias doenças podem ser causadas pela Y. enterocolitica em humanos, desde uma diarréia moderada até complicações mais graves como, adenite mesentérica e artrite reumatóide. Com o objetivo de determinar a ocorrência e o isolamento desse agente na linha de abate de suínos, foram coletadas amostras de tecidos biológicos, excrementos e suabes de carcaças de suínos ao abate e de facas  em um matadouro-frigorífico supervisionado pelo Serviço de Inspeção Estadual do estado de São Paulo (SISP)

Avaliação do consumo de leite e produtos lácteos informais e do conhecimento da população sobre os seus agravos à saúde pública, em um município do Estado de São Paulo, Brasil / Evaluation of consumption of milk and its informal derivatives and the knowledge of the population about the public health problems in a municipal district in São Paulo, Brazil

O leite é um alimento de elevado valor nutricional e considerado um dos principais produtos de origem animal consumidos nos dias de hoje. O Brasil destaca-se como um dos principais produtores de leite do mundo e desde 2006 vem apresentando aumento crescente em sua produção. Por outro lado, também se observa desde o início da década de 1990, crescimento do chamado leite e produtos lácteos informais (alimentos comercializados sem qualquer tipo de inspeção), representando desafio a ser vencido com relação à segurança alimentar. Muitas são as enfermidades que podem ser transmitidas por esses alimentos, como por exemplo, a tuberculose e a brucelose e poucos são os consumidores que estão atentos a essas informações. O fato agrava-se ao pensar que o leite e os produtos lácteos compõem a dieta de muitas crianças, idosos e pessoas imunocomprometidas, o que pode tornar esse quadro mais grave. O presente trabalho teve como objetivo avaliar, por meio da aplicação de questionários em 23 bairros do município de São José do Rio Preto, o perfil de 805 entrevistados, que consomem ou não leite e produtos lácteos, o conhecimento que os mesmos apresentavam sobre as doenças transmitidas pelo leite e seus derivados e o significado dos selos dos Serviços de Inspeção. Como resultado, observou-se que 97,76% (787/805) dos entrevistados consomem leite ou seus derivados, e aproximadamente 30% são obtidos por meios informais de comércio. A forma de consumo desse produto é bastante variável, mas 3,93% (31/787) dos consumidores do município ainda ingerem leite cru sem a fervura ou pasteurização e 65,96% (531/805) dos consumidores desconhecem a possível transmissão de doenças através da ingestão do leite. Além disso, 80,24% (646/805) dos consumidores não procuram nas embalagens se o produto passou por algum tipo de fiscalização antes de chegar ao comércio - identificação através dos selos dos órgãos de inspeção sanitária. A obtenção desses dados visou...(AU)

Milk is a food of high nutritional value and is considered one of the main animal products consumed today. At the same time, Brazil stands out as one of the leading milk producers in the world and, since 2006 has been showing growing increase in its production. On the other hand, it is observed from the beginning of the 1990s, the growth of so-called informal milk and milk products (foods marketed without any kind of inspection), representing a challenge to be overcome related to food security. There are many diseases that can be transmitted by such foods, as tuberculosis and brucellosis, and there are few consumers who are aware of such information. The fact worsens the thought that milk and dairy products make up the diet of many children, elderly and weakened persons, which may make it more severe. Thus, this study aims to evaluate, through the use of questionnaires in 23 districts of São José do Rio Preto, the profile of 805 consumers or not, of milk and dairy products, the knowledge that they have about the diseases transmitted by milk and dairy products and the significance of the seals from Inspection Services. As a result, it was found that 97.76% (787/805) respondents consume milk or its derivatives, and about 30% are obtained by informal means of commerce. The form of consumption of this product is quite variable, but the work shows that 3.93% (31/787) of the consumers still ingest raw milk without boiling or pasteurization and 65.96% (531/805) of consumers are unaware of the possible transmission of disease by the ingestion of milk. Furthermore 80.24% (646/805) of consumers do not seek the packaging if the product has gone through some kind of supervision before reaching the trade - identification through stamps organs of sanitary inspection. Obtaining such data aims to collaborate in future for the development of prevent programs in the city, and the information...(AU)

DECREASE OF SOMATIC CELL COUNT IN RAW MILK THROUGH EXPERIMENTAL MODEL OF FRAUD / Diminuição da contagem de células somáticas em leite cru através de modelo experimental de fraude.

Tendo em vista busca por rendimento industrial e lucratividade, as indústrias propuseram pagamento pela qualidade do leite produzido, assim o produtor receberia bonificação ou penalização referentes à composição do leite como teores de gordura, proteína e contagem de células somáticas (CCS). Porém, quando existem programas de melhoria de qualidade onde o produtor poderá ser bonificado/penalizado ocorre o risco de fraudes surgirem, a fim de atender o estabelecido. O objetivo desse estudo foi verificar a viabilidade do modelo experimental de fraude mediante a diminuição da CCS pela subtração de porções de sobrenadante de leite mantido em estação e avaliar as consequências desta subtração sobre alguns componentes do leite. Para tal, foram utilizadas amostras de leite de 18 animais, submetidas a dois diferentes períodos de estação e/ou repouso (30 e 60 minutos) e posteriormente subtraídos 5,0%, 7,5% e 10,0% dos sobrenadantes do leite oriundos destas vacas. Os resultados demonstraram que é possível diminuir a CCS do leite por meio do modelo experimental de fra