Detection of nontuberculous mycobacteria from water buffalo raw milk in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

On line extraction and separation of bendiocarb, methomyl, methylparathion, and pentachlorophenol pesticides from raw milk

A high performance liquid chromatography (HPLC) method for extraction and determination of pesticides from raw milk was developed. The method involves direct injection of raw milk samples on a bovine serum albumin-dimethyl-octyl-silica gel (BSA-Si-Cs) column. The mobile phase 0.05 mol.L-1 phosphate buffer pH6.0 in acetonitrile (70:30 v/v) was employed for extraction and separation of bendiocarb, methylparathion, pentachlorophenol, and methomyl pesticides. The method shows good results of recovery in the pesticides studied, higher than 99.6%.

Avaliação de métodos de análises para determinação da contagem de células somáticas no leite cru, mantido em tanque de resfriamento

We analyzed the quality of raw milk from eight dairy farms in Rio Grande do Norte stored in a cooling tank , in order to evaluate methods for determining somatic cell counts (SCC). The Somaticell® kit and a portable Direct Cell Counter (DCC) were compared with each other and with the MilkoScanTM FT+ (FOSS Denmark), which uses Fourier Transform Infrared (FTIR) spectroscopy). Direct cell counter data were processed for somatic cell scores (log-transformed somatic cell count) and analyzed with the SAS®, statistical package , Statistical Analysis System, (SAS, INSTITUTE, 1998). Comparison of means and correlation of somatic cell scores were conducted using Pearson s correlation coefficient and the Tukey Test at 1 %. No significant difference was observed for comparison of means. The correlation between somatic cell scores was significant, that is, 0.907 and 0.876 between the MilkoScanTM FT+ and the Somaticell® kit and Direct Cell Count (DCC) respectively, and 0.943 between the Somaticell® kit and Direct Cell Count (DCC). The methods can be recommended for monitoring the quality of raw milk kept in a cooling tank in the production unit

Digital Light Microscopy of Macroporous Ceramics Processed by Consolidation Casting with Mixtures of Native Starches

Porosity in starch consolidation casting technique is rightly related to original size and morphology of starch granules, leaving a pore structure after burning out. This work reports the results for the addition of different native potato and corn starch proportions in suspension,; with TiO(2) (rutile) powder. Gelling temperature have been defined after observation under light microscopy using a heating stage. Analysis of porous network and isolated pores have been clone from images of samples surfaces obtained by depth from focus reconstruction, revealing a qualitative correlation of pores characteristics and starches additions in suspensions, suggesting that the presence of isolated or interconnected pores can be handled by starches selection to control the amylopectin and amylose contents in slurries. Also, the analysis of porous fraction distribution shows no consistent pattern through specimens' volume according to starches in mixtures.

Evaluation of stitched fabric composite processed by RTM in quasi-static test

Stitched fabrics have been widely studied for potential application in aircraft structures since stitch yarns offer improvements in the out-of-plane mechanical properties and also can save time in the lay up process. The down side of stitch yarns came up in the manufacturing process of fabric in which defects introduced by the needle movement creating fiber-free-zones, fiber breakage and misalignment of fibers. The dry stitched carbon fabric preform has mainly been used in the Resin Transfer Molding (RTM) process which high fiber content is aimed, those defects influence negatively the injection behavior reducing the mechanical properties of final material. The purpose of this research work focused on testing in quasi-static mechanical mode (in-plane tension) of a monocomponent resin CYCOM (R) 890 RTM/carbon fiber anti-symmetric quadriaxial fabric stitched by PE 80Dtex yarn processed by RTM. The evaluation consisted in comparing the scatter of the quasi-static test with the attenuation of ultrasonic maps, which show the path of the resin and possible dry spots considering that interference of yarn in resin flow is detectable in ultrasonic measurement. Microscopic analysis was also considered for further evaluation in case of premature failure. (C) 2011 Published by Elsevier Ltd. Selection and peer-review under responsibility of ICM11

Orientation dependence of stored energy of cold work in semi-processed electrical steels after temper rolling

Microhardness measurements were carried out in a low carbon lamination steel after 6% of temper rolling, in order to evaluate local variations of work hardening as a function of crystallographic orientation. EBSD (electron back scattered diffraction) was used to determine grain orientations with respect to individual rolling planes and rolling directions. Hardness was shown to increase with the local Taylor factor. TEM observations and a well-known dislocation hardening model were used to confirm the equivalence between hardness values and the stored energy of cold work. A definite correlation between stored energy and Taylor factors could therefore be established, being more consistent than previous data reported in the literature. The improvement was thought to be related to the rather small plastic deformation, during which Taylor factors could be considered to remain constant. (c) 2006 Elsevier B.V. All rights reserved.

Time-dependent alterations of soluble and cellular components in human milk

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation of brucella spp from milk of brucellosis positive cows in São Paulo and Minas Gerais states

A brucelose é uma zoonose crônica de importância para a Saúde Pública. Considerando o pequeno número de dados brasileiros sobre a sua presença em leite cru e derivados não-pasteurizados, estudamos a presença de brucelas em leite de animais sorologicamente positivos. A soroaglutinação rápida (SAR), a soroaglutinação lenta (SAL) e a soroaglutinação lenta com tratamento do soro com 2-mercaptoetanol foram utilizados para identificar os animais positivos nas propriedades estudadas. Amostras diárias de 300 ml de leite foram colhidas por três dias de todos os quartos mamários produtivos (75 ml/teto). As amostras eram misturadas e centrifugadas. Parte do sedimento e do sobrenadante foi inoculada em meios de Farrel e Brodie-Sinton (BS) suplementados com agentes antimicrobianos. As placas e tubos foram cultivados por sete dias a 37ºC, em microaerofilia. As colônias suspeitas no meio BS foram imediatamente repicadas para ágar-Brucella, e cultivadas sob a mesma condição. Os microrganismos isolados foram submetidos a procedimentos de identificação, incluindo a coloração de Gram, requerimento de CO2, produção de H2S, atividade da urease e crescimento na presença de tionina e fucsina. Das 49 amostras examinadas, isolou-se Brucella abortus de 15 (30,61%). Os biótipos isolados foram: biótipo 1 em uma amostra (2,04%), biótipo 2 em oito (16,32%) e biótipo 3 em seis amostras (12,25%)

Somaticell® as a screening method for somatic cell count from bovine milk

Os objetivos do presente estudo foram avaliar a correlação entre a contagem eletrônica de células somáticas (eCCS) com o Somaticell® sob diferentes níveis de contagem de células somáticas (CCS) do leite e patógenos causadores de mastites, além de calcular a sensibilidade, especificidade e valores preditivos do Somaticell® utilizando diferentes limites de CCS estabelecidos pelos diferentes países. Trezentos e quarenta amostras de leite foram coletadas assepticamente após realização do California Mastitis Test (CMT). O Somaticell® e a eCCS foram realizados em todas as amostras de leite. A correlação entre o Somaticell® e a contagem eletrônica foi determinada de acordo com o CMT, patógeno isolado e escore de eCCS. de acordo com os escores de CCS estabelecidos, 26,5% das amostras de leite apresentaram escore 1 (69-166 x10³células mL-1), 26,8% escore 2 (167-418x10³células mL-1), 27,4% escore 3 (419-760x10³células mL-1) e 19,4% escore 4 (761 to 1970x10³células mL-1). A eCCS e o Somaticell® apresentaram correlação positiva em quase todos os escores estudados (exceto escore 2 e 3). O valor de r obtido entre CCS e o Somaticell® foi de 0,32. Observou-se que, quando o limite de CCS estabelecido aumentou, a sensibilidade decresceu e os valores de especificidade aumentaram. Os valores preditivos apresentaram-se constantes em todos os limites. Quando o limite de CCS era baixo (<760,000 células mL-1), Somaticell® forneceu resultados consistentemente mais elevados que os valores de CCS. Já para amostras com CCS elevada, Somaticell® resultou em menores contagens que a eCCS. A correlação entre os dois métodos permaneceu relativamente constante em todas as condições e os valores de sensibilidade e especificidade do teste são altamente dependentes do limite estabelecido. Os resultados deste trabalho sugerem que o Somaticell® não é útil para avaliar a CCS do leite, pois seus resultados são significativamente diferentes da eCCS, no entanto, pode ser utilizado como método de triagem, tal como o CMT, para a detecção do aumento da CCS do leite.

Molecular identification and thermoresistance to boiling of Nocardia farcinica and Nocardia cyriacigeorgica from bovine bulk tank milk

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Breed and season influence on milk quality parameters and in mastitis occurrence

Os objetivos do presente estudo foram avaliar a performance das raças Jersey e Holandesa em diferentes condições de pluviosidade (estações seca e chuvosa) através do monitoramento de aspectos relacionados com a mastite subclínica (contagem de células somáticas, isolamento microbiológico, tipo de patógeno isolado), qualidade do leite (lactose, proteína, gordura e sólidos totais) e produção (media da produção de leite) de ambas as raças. O estudo foi conduzido em propriedade leiteira localizada no estado de São Paulo, Brasil. Oito visitas foram realizadas a propriedade, quatro em período de alta pluviosidade e outro em período de baixa pluviosidade. Amostras de leite foram colhidas de todos os animais para contagem eletrônica de células somáticas e determinação dos principais componentes do leite (proteína, gordura, sólidos totais, lactose). Gordura, proteína, sólidos totais e produção leiteira foram influenciadas tanto pela raça como pela estação, apresentando tendências similares para ambas raças em ambas estações. A contagem de células somáticas (CCS) apresentou resultados similares para ambas as raças. Vacas holandesas com infecção intramamária (IIM) apresentaram um maior aumento na CCS quando comparadas as vacas Jersey (P<0.001). Na estação seca, 53 animais tiveram IIM em pelo menos um mês do estudo. Destes, 32 eram da raça Holandesa e 21 da raça Jersey. Na estação chuvosa, 65 animais possuíram infecção intramamária, dos quais 43 eram da raça Holandesa e 22 da raça Jersey. A frequência de casos de IIM foi maior na estação chuvosa em comparação à estação seca. Vacas Jersey apresentaram uma menor chance de desenvolver sinais e sintomas de IIM em relação às vacas holandesas na estação chuvosa (razão de chances=0.52). O maior numero de casos de IIM na estação chuvosa pode ter contribuído para uma menor taxa de lactose no leite para ambas as raças, sendo que esta pode ser considerada como um indicador do status de IIM. Existiu uma prevalência de patógenos contagiosos durante todo o experimento. O modelo estatístico aplicado mostrou que patógenos ambientais foram mais frequentemente isolados da raça Jersey, independentemente do período do estudo. Aparentemente existem diferenças na resposta imune das raças Jersey e Holandesa.

Molecular detection of Streptococcus agalactiae in bovine raw milk samples obtained directly from bulk tanks

Mastitis is the most common infectious disease affecting dairy cattle; in addition, it remains the most economically important disease of dairy industries around the world. Streptococcus agalactiae, a contagious pathogen associated with subclinical mastitis, is highly infectious. This bacterium can cause an increase in bulk tank bacterial counts (BTBC) and bulk tank somatic cell counts (BTSCC). The microbiological identification of S. agalactiae in samples from bulk tanks is an auxiliary method to control contagious mastitis. Thus, there are some limitations for time-consuming cultures or identification methods and additional concerns about the conservation and transport of samples. Bulk tank samples from 247 dairy farms were cultured and compared through polymerase chain reaction (PCR), directed to 16S rRNA genes of S. agalactiae, followed by BTBC and S. agalactiae isolation. The mean value of BTBC was 1.08 x 10(6) CFU mL(-1) and the bacterium was identified through the microbiological method in 98 (39.7%; CI95% = 33.8-45.9%) and through PCR in 110 (44.5%; CI95% = 38.5-50.8%) samples. Results indicated sensitivity of 0.8571 +/- 0.0353 (CI95% = 0.7719-0.9196) and specificity of 0.8255 +/- 0.0311 (CI95% = 0.7549-0.8827). The lack of significant difference between microbiological and molecular results (kappa = 0.6686 +/- 0.0477 and CI95% = 0.5752-0.7620) indicated substantial agreement between the methods. This suggests that PCR can be used for bulk tank samples to detect contagious mastitis caused by S. agalactiae. (C) 2011 Elsevier Ltd. All rights reserved.

Detection of Toxoplasma gondii in the milk of experimentally infected Wistar female rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on Prototheca zopfii strains isolated from bovine milk

Prototheca zopfii has been considered one of the most important causes of environmental mastitis in Brazil. These algae are refractory to conventional therapy and cause great damage to the mammary gland. The present study evaluated the in vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on 27 P. zopfii strains isolated from the milk of cattle. Low concentrations of sodium hypochlorite (0.0390625-0.15625%) and iodine (0.15625-0.625%) were effective against the isolates. These antiseptics may be recommended for hygiene routines, pre and postdipping and cauterization of bovine mammary glands infected by P. zopfii. (C) 2009 Elsevier Ltd. All rights reserved.

Microbiological evaluation of milk samples positive to California Mastitis Test in dairy buffalo cows (Buballus bubalis)

In order to observe the microbiological status of CMT positive samples, 734 apparently health mammary quarters from buffalo cows were submitted to physical evaluation, strip cup test and CMT. After milk samples inoculation in 10% ovine blood agar base media and in MacConkey agar and incubation under aerobic condition for 72 hours at 37 degrees C, identification was proceeded. According to CMT, 227 quarters (30,93%) were positive, among them 73 (32,16%) presented 1+ reaction, 53 (23,35%) were 2+ and 101 (44,49%) were 3+. Microbiological exams of such samples were positive in 147 (64,76%) out of 227 CMT positive samples and among the remaining 72 (31,72%) were negative and 8 (3,52) were contaminated. In the 147 microbiological positive samples 204 bacteria were found in pure or associated growth and the most frequent agents were: Corynebacterium sp (59,25%); Staphylococcus sp (17,65%) among which 86,11% were coagulase negative and 13,89% were coagulase positive; and Micrococcus sp (6,37%). The results revealed that, excluding the eight contaminated samples, 147 (67,12%) quarters out of 219 CMT positive could be considered as bacteria-carrier and that even in a smaller percentage false-positive results can cause problems in a sanitary program for mastitis control in dairy buffalo cows.