AN IMPROVED ENRICHMENT PROCEDURE FOR THE ISOLATION OF YERSINIA-ENTEROCOLITICA AND RELATED SPECIES FROM MILK

A new enrichment procedure is proposed to improve the isolation of Yersinia enterocolitica and related species from milk. This procedure uses tryptic soy broth plus Polymyxin (5 IU/ml) and Novobiocin (10 mug/ml) - TSPN broth - incubated at 18-degrees-C for 3 d. Using raw milk and pasteurized milk inoculated with Yersinia strains, the efficiency of this procedure was compared to that of SB broth (sorbitol bile salts broth) incubated at 4-degrees-C for up to 21 d. Despite of the presence of antibiotics in TSPN broth, there were difficulties in recovering Yersinia organisms. Nevertheless, TSPN broth incubated at 18-degrees-C for 3 d showed better efficiency than that other method. In pasteurized milk samples, TSPN medium at 18-degrees-C for 3 d gave better results than the SB broth at 4-degrees-C for 7 d, showing that the proposed procedure is the preferable one due to the shorter period of incubation.

Evaluation of different rhodium modifiers and coatings on the simultaneous determination of As, Bi, Pb, Sb, Se and of Co, Cr, Cu, Fe, Mn in milk by electrothermal atomic absorption spectrometry

Different kinds of modifiers and coatings on the integrated platform of transversely heated graphite atomizer (THGA) have been tested for the simultaneous determination of two group of elements: the first, the more volatile, formed by arsenic, bismuth, lead, antimony and selenium; the second, the less volatile, formed by cobalt, chromium, cupper, iron and manganese in milk by electrothermal atomic absorption spectrometry. Different Rh-modifiers were studied, such as Rh-coated platforms (Rh), carbide plus rhodium coated platforms (W-Rh, Zr-Rh), carbide-coated platforms (W and Zr) with co-injection of RhCl3, solutions and uncoated platforms with injection of solutions of Pd(NO3)(2), Mg(NO3)(2), and RhCl3. Milk samples were diluted 1:10 in 1.0% HNO3 and injected into the tube. The mass of modifier deposited and co-injected in the tube and the use of end capped tubes were also evaluated in order to improve the electrothermal behavior of analytes. Integrated platform pretreated with W plus co-injection RhCl3 for first group and pretreated with W-Rh for second group were elected. For 20 mu L injected samples the analytical curves in the 5.0-20.0 mu g L-1 concentration range have good linear correlation coefficients (r > 0.998). Relative standard deviations (n = 12) are < 6% and the calculated characteristic masses are between 5 pg and 62 pg.

Grazing time and milk production of crossbred cows in a rotational area of Elephant grass and Tanzania grass

This study was carried out in January of two consecutive years. The objective was to evaluate the grazing time, the grazing rate and the milk production of crossbred cows, managed in two rotational grazing areas, one with elephant grass (Pennisetum purpureum Schum. cv. Guaçu) and another one with Tanzania grass (Panicum maximum Jacq. cv. Tanzania), both with natural shade in the rest area. The experiment was divided in two phases, the first with 12 cows that remained day and night in the paddock and were milked twice a day. In the second phase 15 cows were observed and remained 10.43 hours in the paddock during the day, and were kept in a corral during the night. Each cow was observed every 15 minutes. Grass specie had no effect on milk production in the two phases of the experiment. The grazing time was 564 and 474 minutes and the grazing rate was 28.7 and 24.4 minutes/hour for the Elephant grass and Tanzania grass, respectively, in first phase. In the second phase,the grazing time was 461 and 426 minutes and the grazing rate was 42.7 and 39.4 minutes/hour for the Elephant grass and Tanzania grass, respectively. Correlations were observed (P<0.01) between grazing rate and the minimum temperature (-0.68), the maximum temperature (-0.76), the relative humidity (0.44) and the THI (-0.76).

Effect of shade and water sprinkling on physiological responses and milk yields of Holstein cows in a semi-arid region

The effects of shading and water sprinkling on physiological responses and milk production were studied in Holstein cows managed in an equatorial semi-arid region. Five cows were observed for 36 days during May and June 2009. We studied the effects of sun, shade, shade plus single sprinkling, shade plus sprinkling every 2. h and shade plus sprinkling every hour on physiological responses, i.e., rectal temperature, coat surface temperature and respiratory rate, and on milk production. Environmental variables, i.e., air temperature, black globe temperature, mean radiant temperature, relative humidity and wind speed, were recorded every 30. min throughout the observation period. The statistical analysis utilized a 5×5 replicated latin square, and the results showed reduced values in physiological variables in cows that were protected from short wave radiation and that received water sprinkling; milk yield increased by approximately 3.5. kg in such animals relative to those not receiving such treatment. However, the milk yield for the morning milking presented similar mean values regardless of whether the animals received shading and sprinkling benefits, which indicates that frequency of sprinkling and amount of time (permanence) in the shade were critical for the effect on milk yield. © 2013 Elsevier B.V.

Nutrients balances and milk fatty acid profile of mid lactation dairy cows supplemented with unsaturated fatty acid

The objective was to evaluate the effect of unsaturated fatty acid sources supplementation on nutrients balances and milk fatty acid profile of mid lactation dairy cows. Twelve Brazilian Holstein cows in the mid lactation (mean of 128 days) and (580 ± 20kg of weight; mean ± SD) with milk yield of 25kg/d were assigned randomly into three 4 × 4 Latin square, fed the following diets: control (C); refined soybean oil; (SO); whole soybean raw (WS) and; calcium salts of unsaturated fatty acids (CSFA). Milk yield was 26.6; 26.4; 24.1 and 25.7 to the diets CO, SO, WS and CSFA respectively. Cows fed the WS treatment produced less milk (1.95kg/d of milk), fat and lactose than did cows fed the SO and CSFA. Cows fed the CSFA treatment showed less blood, urine (g/d) concentrations of N more energetic efficiency and intake of energy than did cows fed the SO treatment. Cows fed the unsaturated fatty acids sources showed more C18:2 cis-9, trans-11 CLA and trans-C18:1 FA concentration in milk than did cows fed the CO treatment. Diets with whole soybeans and soybeans oil provide more efficient digestive processes, and increase milk composition of unsaturated fatty acids.

Molecular characterization and clonal diversity of methicillin-susceptible Staphylococcus aureus in milk of cows with mastitis in Brazil

Mastitis is an important disease for the dairy industry worldwide, causing economic losses and reducing milk quality and production. Staphylococcus aureus is a worldwide agent of this intramammary infection, which also causes foodborne diseases. The objective of this study was to determine the frequency of methicillin-susceptible Staphylococcus aureus (MSSA) isolates in milk of mastitis cows in Brazil and to analyze the genetic lineages and the content of antimicrobial resistance genes and virulence factors among these isolates. Fifty-six MSSA isolates were recovered from 1,484 milk samples (positive for the California mastitis test) of 518 cows from 11 different farms in Brazil (representing 51% of total Staph. aureus obtained), and they were further characterized. Methicillin-susceptible Staphylococcus aureus were isolated from 3.7% of California mastitis test-positive tested milk samples and from 6.2% of tested mastitic cows. Methicillin-susceptible Staphylococcus aureus isolates were characterized by spa typing, agr typing, and multilocus sequence typing, and resistance and virulence traits were investigated by PCR. Seven spa types were identified among MSSA (% of isolates): t127 (44.6), t605 (37.5), t002, t1784, t2066 (1.8), and 2 new ones: t10856 (10.7) and t10852 (1.8). Five distinct sequence types (ST) were detected (% of isolates): ST1 (46.4), ST126 (37.5), ST133 (10.7), ST5 (3.6), and a novel ST registered as ST2493 (1.8). Resistances were detected for streptomycin, chloramphenicol, and tetracycline. One strain contained the chloramphenicol resistance gene (fexA; included within transposon Tn558) and 3 strains contained the tetracycline resistance gene [tet(K)]. Methicillin-susceptible Staphylococcus aureus strains were susceptible to most of the antibiotics studied and lacked the virulence genes of Panton-Valentine leukocidin (lukF/S-PV), toxic shock syndrome toxin 1 (tst), exfoliative toxin A (eta), and exfoliative toxin B (etb), as well as the genes of the immune evasion cluster. Methicillin-susceptible Staphylococcus aureus isolates were detected in a relatively low proportion of cows with mastitis (6.2%) and recovered isolates presented high diversity of genetic lineages, with CC1 and CC126 the predominant clonal complexes, and CC133 also being detected. Larger epidemiological studies with molecular characterization of isolates are required to deepen the knowledge on the circulating genetic lineages among the cow population with mastitis. © 2013 American Dairy Science Association.

Occurrence of heat waves in Triangulo Mineiro and Alto Paranaiba and their effect on milk yield and feed intake in cattle

The aim of this study was to estimate milk production and food consumption during the occurrence of heat waves in the Triangulo Mineiro and Alto Paranaiba, MG by means of bioclimatic zoning based on the Temperature and Humidity Index (THI). Therefore a history of heat wave occurrence between the years 2000-2010 was compiled. The decline in milk production (DMP) and reduced food consumption (RFC) were simulated in cities where periods of heat waves were identified. Frutal and Ituiutaba had the highest rate of heat wave occurrence per year. The DMP and RFC showed bioclimatic differences between the cities of Uberaba, Ituiutaba and Frutal. The cities with the best bioclimatic conditions were Sacramento and Patrocinio, as they presented a THI classified outside of the emergency range, with a night THI of below 76 and without heat waves. Therefore, the occurrence of heat waves can impair food intake and decrease milk production, thereby most effectively demonstrating the effects of thermal stress on dairy cows in the Triangulo Mineiro and Alto Paranaiba, MG region.

Molecular epidemiology of Staphylococcus aureus isolates at different sites in the milk producing dairy farms

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

Ocorrência de Bacillus cereus em leite integral e capacidade enterotoxigênica das cepas isoladas

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.

Suckling and allosuckling in river buffalo calves and its relation with weight gain

The aim of the present study was to investigate if river buffalo calves (Bubalus bubalis) have equal access to all cows and if milk is thus equally available to all of them. We recorded suckling/allosuckling behaviour and weight gain (WG) of 29 calves (14 males and 15 females), with special consideration to their sex, birth order (BO) and age. Cows' nursing behaviour and milli production (MP) were also considered. While males tended to be born earlier than females during this study, this was not the trend in the overall herd records. The cows' MP was not effected by the calves' sex. However, bull-calves presented greater mean WG, and mean times spent in individual filial (IF) and in communal nonfilial (CNF) suckling than heifer-calves, which showed greater communal filial (CF) suckling than the former during the first 4 months of life. The WG was associated with IF for bull-calves (r = 0.680 and 0.765, respectively, for the periods from birth to 4th and 8th months of age), and to CNF for heifer-calves (r = 0.628, for the period from birth to 8th month). Results from multiple regression analysis showed independent effects of each suckling category on the calf WG, and such effects were variable according to the calf's sex. BO was negatively correlated to calves' WG (bull-calves: r(s) = - 0.873 and - 0.799, from birth to 4th and gth months, respectively; heifer-calves: r(s) = - 0.531 from birth to 4th month). Specifically for bull-calves, there was a positive correlation between BO and MP (r(s) = 0.528 and 0.633, from birth to 4th and 8th months of age, respectively). The correlation between BO and IF was negative in both sexes, indicating that calves that were born early had more opportunities to suckle individually from their mothers. For heifer-calves, BO was positively correlated with CF (two periods), and negatively with CNF (from birth to 8th month of age), suggesting that heifer-calves were most often accompanied by other calves during suckling when they were born later. The data taken together indicate that sex and/or BO influenced decisively social interactions during suckling, promoting differential development among the calves. In animal husbandry, if a homogenous WG is desired, these factors have to be taken into consideration. (C) 2000 Elsevier B.V. B.V. All rights reserved.

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.

Analysis of the electrical conductivity in milking fractions as a mean for detecting and characterizing mastitis in goats

The aim of the work was to study the effect of milking fraction on electrical conductivity of milk (EC) to improve its use in dairy goat mastitis detection using automatic EC measurements during milking. The experiment was carried out on a group of 84 Murciano-Granadina goats (28 primiparous and 56 multiparous). Goats were in the fourth month of lactation. A linear mixed model was used to analyse the relationship between EC or somatic cell count (SCC) of gland milk and parity, mammary gland health status, analysed fraction (first 100 mL=F-1; machine milk=F-2; and stripping milk=F-3) and their first order interactions. Additionally, the mastitis detection characteristics (sensitivity, specificity, positive predictive value and negative predictive value) of SCC and EC were studied at different thresholds.All factors considered were significant for EC and SCC. EC decreased significantly as milking progressed (from F-1 to F-3) in both healthy and infected glands. EC was not significantly different between healthy and infected glands in F-1 and F-2 fractions, but EC of healthy glands (5.01 mS/cm) was significantly lower than in infected glands (5.03 mS/cm) at F-3.Mastitis detection characteristics of EC did not differ amongst studied fractions. The small significant difference of EC between healthy and infected glands obtained in F-3 fraction did not yield better sensitivity results compared to F-1 and F-2. The best EC mastitis detection characteristics were obtained at 5.20 mS/cm threshold (sensitivity of 70% and specificity of 50%). The best SCC mastitis detection characteristics were obtained at 300,000 cells/mL threshold and F-3 fraction (sensitivity of 85% and specificity of 65%).It was concluded that mastitis detection characteristics of EC were similar in the three milking fractions analysed, being slightly better for SCC in F-3 fraction. As shown in previous studies, there are no factors other than the mammary gland health status that affect milk EC and should be considered in the algorithms for mastitis detection to improve the results. (C) 2012 Elsevier B.V. All rights reserved.

Voluntary intake of dry matter and performance of Nellore cows and their Nellore and crossbred Simental x Nellore calves

This study aimed to investigate the influence of calf genetic group on the performance of Nellore lactating cows. The variables studied included milk ingestion, pasture intake, calf weight gain and cow body weight. A total of 13 Nellore calves and 8 crossbred Simental x Nellore calves were used, all born from Nellore mothers. During the experimental period of 210 days, calves were weighted at birth and at 30-day intervals, whereas cows were weighed every 14 days. Milk intake was estimated using the weigh-suckle-weigh method. Pasture intake was determined 6, 12, and 24 weeks after birth by using chromic oxide as an external marker and indigestible acid detergent fiber as an internal marker. Average weight of crossbred calves was 10.1 kg and their milk intake was 0.42 kg higher than in purebred calves. There was no difference, however, in pasture intake between genetic groups. In Nellore cows, body weight and pasture consumption (9.2 kg of DM/cow/day) were not influenced by calf heterosis.

Effects of calcium salts of polyunsaturated fatty acids on productive and reproductive parameters of lactating Holstein cows

Two experiments evaluated milk production, serum progesterone and insulin, and reproductive performance of lactating Holstein cows receiving or not receiving Ca salts of polyunsaturated fatty acids (PUFA), or receiving Ca salts of PUFA at different daily frequencies. In experiment 1, 1,125 cows randomly distributed in 10 freestall barns were enrolled. Barns were assigned randomly to receive a high-concentrate diet containing (PI?) or not containing (control, CON) 1.1% (dry matter basis) Ca salts of PUPA. Diets were offered 6 times daily, whereas the Ca salts of PUFA were included in the PF treatment in the first feeding of the day. In experiment 2, 1,572 cows were randomly distributed in 10 freestall barns, which were assigned randomly to receive a diet similar to PF, but with Ca salts of PUFA included only in the first feeding of the day (PF1X), or equally distributed across all 6 feedings (PF6X). During both experiments, cows were artificially inseminated 12 h after the onset of estrus. Once per month, cows that did not conceive to artificial insemination were assigned to a fixed-time embryo transfer protocol. Pregnancy was determined via transrectal ultrasonography 28 and 60 d after expected ovulation. Pregnancy loss was considered in cows that were pregnant on d 28 but nonpregnant on d 60. During both experiments, feed intake, milk yield, and milk protein and fat content were recorded weekly. Blood samples were collected concurrently with embryo transfer. During experiment 1, feed intake was similar between treatments. Compared with CON, PF cows had greater milk yield (37.8 vs. 35.3 kg/d), and reduced milk fat content (3.41 vs. 3.55%). However, PF cows had reduced pregnancy losses per service compared with CON (12.6 vs. 18.3%). Serum progesterone was greater and serum insulin tended to be greater in primiparous cows receiving PF compared with CON cohorts (4.50 vs. 3.67 ng of progesterone/mL, and 10.4 vs. 7.5 mu UI of insulin/mL). During experiment 2, no treatment effects were detected for feed intake, milk yield, or milk fat, whereas PF1X cows tended to have reduced pregnancy losses per service compared with PF6X (14.4 vs. 18.4%). In summary, feeding Ca salts of PUFA to dairy cows increased milk production, did not alter feed intake, and reduced pregnancy losses per service. Further, the total daily amount of Ca salts of RITA should be fed during the first feeding of the day to optimize its benefits on pregnancy maintenance of dairy cows.

Voluntary intake of dry matter and performance of Nellore cows and their Nellore and crossbred Simental × Nellore calves

This study aimed to investigate the influence of calf genetic group on the performance of Nellore lactating cows. The variables studied included milk ingestion, pasture intake, calf weight gain and cow body weight. A total of 13 Nellore calves and 8 crossbred Simental × Nellore calves were used, all born from Nellore mothers. During the experimental period of 210 days, calves were weighted at birth and at 30-day intervals, whereas cows were weighed every 14 days. Milk intake was estimated using the weigh-suckle-weigh method. Pasture intake was determined 6, 12, and 24 weeks after birth by using chromic oxide as an external marker and indigestible acid detergent fiber as an internal marker. Average weight of crossbred calves was 10.1 kg and their milk intake was 0.42 kg higher than in purebred calves. There was no difference, however, in pasture intake between genetic groups. In Nellore cows, body weight and pasture consumption (9.2 kg of DM/cow/day) were not influenced by calf heterosis.