J. Dairy Sci. 873561–3573 American Dairy Science Association, 2004. Guidelines for Monito
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J.Dairy Sci.90:1122–1132©American Dairy Science Association,2007.Evaluating Mid-infrared Spectroscopy as a New Technique for Predicting Sensory Texture Attributes of Processed CheeseC.C.Fagan,*1C.Everard,*C.P.O’Donnell,*G.Downey,†E.M.Sheehan,‡C.M.Delahunty,§andD.J.O’Callaghan ʈ*Biosystems Engineering,UCD School of Agriculture,Food Science and Veterinary Medicine,University College Dublin,Earlsfort Terrace,Dublin 2,Ireland†Teagasc,Ashtown Food Research Centre,Dublin 15,Ireland‡Department of Nutritional Sciences,University College Cork,Cork,Ireland§Department of Food Science,University of Otago,PO Box 56,Dunedin 9015,New Zealand ʈTeagasc,Moorepark Food Research Centre,Fermoy,Co.Cork,IrelandABSTRACTThe objective of this study was to investigate the po-tential application of mid-infrared spectroscopy for de-termination of selected sensory attributes in a range of experimentally manufactured processed cheese samples.This study also evaluates mid-infrared spectroscopy against other recently proposed techniques for pre-dicting sensory texture attributes.Processed cheeses (n =32)of varying compositions were manufactured on a pilot scale.After 2and 4wk of storage at 4°C,mid-infrared spectra (640to 4,000cm −1)were recorded and samples were scored on a scale of 0to 100for 9attributes using descriptive sensory analysis.Models were devel-oped by partial least squares regression using raw and pretreated spectra.The mouth-coating and mass-form-ing models were improved by using a reduced spectral range (930to 1,767cm −1).The remaining attributes were most successfully modeled using a combined range (930to 1,767cm −1and 2,839to 4,000cm −1).The root mean square errors of cross-validation for the models were 7.4(firmness;range 65.3),4.6(rubbery;range 41.7),7.1(creamy;range 60.9),5.1(chewy;range 43.3),5.2(mouth-coating;range 37.4),5.3(fragmentable;range 51.0),7.4(melting;range 69.3),and 3.1(mass-forming;range 23.6).These models had a good practical utility.Model accuracy ranged from approximate quantitative predic-tions to excellent predictions (range error ratio =9.6).In general,the models compared favorably with previously reported instrumental texture models and near-infrared models,although the creamy,chewy,and melting models were slightly weaker than the previously reported near-infrared models.We concluded that mid-infrared spec-troscopy could be successfully used for the nondestruc-tive and objective assessment of processed cheese sen-sory quality.Received April 12,2006.Accepted October 30,2006.1Corresponding author:colette.fagan@ucd.ie1122Key words:descriptive sensory analysis,processed cheese,mid-infrared spectroscopy,chemometricsINTRODUCTIONOver 18million tonnes of cheese were produced world-wide in 2004,and processed cheese is an important seg-ment of this market (Wohlfarth and Richarts,2005).The United States,the largest producer of processed cheese (where 20%of all cheese consumed is processed cheese),produced 1,092,000tonnes in 2003(Wohlfarth and Ric-harts,2005).In the same year,the 25countries of the European Union produced 655,000tonnes of processed cheese (Wohlfarth and Richarts,2005).Consumer preference for a food product is principally determined by its sensory characteristics.Accurate mon-itoring and control of sensory properties will facilitate the production of high-quality products.A number of factors determine the final quality and sensory proper-ties of processed cheese (Caric´and Kala ´b,1993).These include the processing conditions used during manufac-ture,the composition of the ingredients,and the propor-tions of those ingredients added to the blend.Sensory profiling allows various quality attributes to be identified and their intensity determined (Brown et al.,2003).Sensory attributes are traditionally assessed by descriptive sensory evaluation using trained panel-ists.However,this is a time-consuming and expensive process that may lack objectivity (Blazquez et al.,2006).Although instrumental techniques such as texture pro-file analysis (TPA )and the 3-point bend test are avail-able for determining the texture attributes of food prod-ucts,these laboratory-based techniques are time-con-suming and require the use of skilled personnel in their execution (Blazquez et al.,2006).Therefore,considerable interest exists in the development of instrumental tech-niques to enable more objective,faster,and less expen-sive assessments of cheese quality to be made,including sensory aspects (Downey et al.,2005).Such a technique would assist producers to maximize yields,increasePREDICTION OF CHEESE SENSORY TEXTURE ATTRIBUTES1123throughput and efficiency,reduce labor costs,and opti-mize product quality,consistency,and customer satisfac-tion.Critical points in the manufacturing process could be monitored to ensure that the final product would meet required specifications.Recently,Kealy (2006)examined cream cheese using TPA,one of the main instrumental techniques for tex-ture measurement,and compared the results with those of a trained taste panel.Although a reasonably strong correlation was found between the taste panel results and TPA-derived hardness and adhesiveness parame-ters,the correlation for cohesiveness was not straightfor-ward.Everard (2005)also investigated the prediction of sensory attributes of processed cheese from instrumen-tal texture attributes derived from TPA,a compression test,and a 3-point bend test.He could predict the texture attributes of firmness,rubbery,creamy,chewy,frag-mentable,and mass-forming with a good level of accu-racy (Everard,2005).Spectroscopic analysis in combination with predictive mathematical models,developed using multivariate data analysis techniques such as partial least squares (PLS )regression,have potential use in controlling and monitoring the quality of raw materials through to the final product in food processing.In particular,infrared spectroscopy has been applied as an objective and nonde-structive technique to provide a rapid and real-time anal-ysis of both composition and quality (Downey,1998;Lefier et al.,2000;Ozen and Mauer,2002;Blazquez et al.,2004).Blazquez et al.(2006)modeled the sensory attributes of processed cheese using near-infrared re-flectance spectroscopy and PLS regression.They found that it was possible to model a number of attributes including firmness,melting,rubbery,and creamy.Two other studies have investigated the prediction of sensory attributes in natural cheese.Downey et al.(2005)and Sørensen and Jepsen (1998)successfully demonstrated that near-infrared spectroscopy in conjunction with PLS regression can be used to predict several sensory attri-butes of Cheddar and Danbo cheeses,respectively.Mid-infrared spectroscopy has been most widely used for de-termination of the fat and protein contents of cheese (Chen and Irudayaraj,1998).Irudayaraj et al.(1999)also investigated the use of mid-infrared spectroscopy to follow texture development in Cheddar cheese during ripening.They demonstrated that springiness could be successfully correlated with a number of bands in the mid-infrared spectra.Research has shown that mid-in-frared spectroscopy is a useful technique for characteriz-ing the changes in proteins during cheese ripening (Ma-zerolles et al.,2001).Pillonel et al.(2003)also found that mid-infrared spectroscopy may be successfully applied to the discrimination of Emmental cheese based on geo-graphic origin.Journal of Dairy Science Vol.90No.3,2007Table 1.Quantity of ingredients (g/kg)used in the production of experimental processed cheese samples Sample Emulsifyingnumber(s)Cheddar Butter Water salt1,10838.70.0161.39.72838.70.0151.619.43,11838.70.0141.929.04,12838.751.6112.99.75,13838.751.6100.019.46,14838.751.690.329.07,15838.7100.061.39.78838.7100.051.619.49,16838.7100.041.929.017,25848.451.6103.29.718,26838.751.6100.019.419,27829.051.6100.029.020,28751.645.2203.29.721,29745.245.2203.219.422,30738.745.2200.025.823,31651.638.7303.216.124,32645.238.7303.222.6No data are currently available on the application of mid-infrared spectroscopy to determine the sensory attributes in processed cheese,or regarding evaluation of mid-infrared spectroscopy in comparison with other technologies in such an application.Therefore,the objec-tives of this study were to investigate the use of mid-infrared spectroscopy in predicting sensory texture attri-butes using a range of experimentally manufactured pro-cessed cheese samples and to compare the models devel-oped with those recently modeled using near-infrared spectra and instrumental texture attributes.These newly presented data allow for the critical evaluation of mid-infrared spectroscopy as a rapid,nondestructive technique for predicting the sensory texture attributes of processed cheese.MATERIALS AND METHODSProcessed Cheese SamplesThirty-two processed cheese batches were manufac-tured in a pilot plant at Moorepark Food Research Cen-tre,Cork,Ireland.The ingredients and formulations are listed in Table 1.The formulations,which were selected to provide samples with compositional ranges that ex-tended beyond those used commercially by processed cheese manufacturers,provided samples with a wide range of sensory characteristics.The ingredients were mixed for 1min in a jacketed cooker (Stephan UMM/SK5Universal cooker;Stephan u So ¨hne GmbH &Co.,Hameln,Germany).The blend was cooked at 80°C for 2min by indirect steam heating.During cooking,the blend was stirred constantly using a knife at 300rpm and a baffle mixer at 80rpm.The cooked blend was stored in food-grade plastic containers (225g capacity),FAGAN ET AL.1124Table2.Vocabulary of sensory attributes,their definitions,and mastication phases used to carry out the sensory analysis of processed cheese samplesSensoryattribute Definition Mastication phaseFirmness The extent of the initial resistance offered by the cheese,Phase1:Judged on thefirst chew using the front teeth ranging from“soft”to“firm”Rubbery The extent to which the cheese returns/springs to its Phase2:Assessed during thefirst2to3chews initial form after biting,ranging from“a little”to“a lot”Creamy The texture associated with cream that has been whipped,ranging from“a little”to“a lot”Chewy The effort needed to break down the structure of the cheese,Phase3:Judged in the middle phase of mastication ranging from“a little”to“a lot”Mouth-coating The extent to which the cheese clings to the insideof the mouth(roof,teeth,tongue,gums),ranging from“a little”to“a lot”Fragmentable Breaks down to smaller versions of itself,ranging from Phase4:Probably judged toward the end of the chewing “a little”to“a lot”Melting The extent to which the cheese melts in the mouth;smooth,velvet fullness in the mouth,ranging from“a little”to“a lot”Mass-forming The extent to which the cheese form a bolus or massin the mouth after chewing,ranging from“a little”to“a lot”Greasy/oily The extent to which a greasy/oily residue is deposited Phase5:Judged at the end of the chewing sequence in the mouth after the cheese is broken down,rangingfrom“a little”to“a lot”which were lidded,allowed to cool,and placed in storage at4°C for4wk.The independent compositional variables for samples1to16were fat and emulsifying salt,and the variables for samples17to32were moisture and emulsifying salt.Descriptive sensory analysis and mid-infrared spectroscopy were carried out at2and4wk postmanufacture.Sensory AnalysisA panel of10assessors(9females and1male),aged 35to55yr old,were selected and recruited in1998and 2000according to international standards(International Organization for Standardization,1993).The panel de-veloped a vocabulary of9texture terms:firmness,rub-bery,creamy,chewy,mouth-coating,fragmentable, melting,mass-forming,and greasy/oily(Table2),which they used to assess each sample.Samples were prepared for analysis in duplicate by removing them from storage and preparing two5-g cubes.These samples were left to equilibrate to room temperature(21°C).Each equili-brated sample was presented to assessors in a glass tumbler covered with a clock glass and labeled with a randomly selected3-digit code.Assessors were provided with deionized water and unsalted crackers to cleanse their palate between tastings.The assessors scored the samples for each attribute by marking on unstructured 100-mm line scales labeled at both ends with extremes of each attribute.The intensity of each of the descriptive terms was recorded using the Compusense v.4.0sensory data acquisition software(Guelph,Ontario,Canada).At each time point,the descriptive sensory analysis took Journal of Dairy Science Vol.90No.3,2007place over2d.The order of tasting was balanced to account for the order of presentation and carryover ef-fects(MacFie et al.,1989).All assessments were con-ducted in individual booths at the sensory laboratory at University College,Cork,which complies with interna-tional standards for the design of test rooms(Interna-tional Organization for Standardization,1988).Mid-Infrared SpectroscopyMid-infrared spectra were collected over the range of 4,000to640cm−1,with a resolution of8cm−1,using an ATI Mattson Infinity Series Fourier transform spectro-photometer(ATI Mattson,Madison,WI)controlled by WinFirst software(ATI Mattson).The sample accessory used for sample presentation was an attenuated total reflectance ZnSe crystal(Graseby Specac Ltd.,Kent, UK),with an incidence angle of45°and6internal reflec-tions.Sixty-four interferograms were coadded before Fourier transformation.Prior to mid-infrared analysis, samples were removed from storage and left to equili-brate to room temperature.This was confirmed prior to analysis using a digital thermocouple(Sensor-Tech Ltd., Co.Louth,Ireland).Processed cheese samples were wiped across the attenuated total reflectance crystal to ensure even and immediate contact.Triplicate spectra were captured for each sample and replicate spectra were averaged prior to data analysis.Multivariate Data AnalysisMultivariate data analysis was carried out using The Unscrambler software(v.8.0;Camo A/S,Oslo,Norway).PREDICTION OF CHEESE SENSORY TEXTURE ATTRIBUTES1125Figure 1.Typical mid-infrared spectrum of processed cheese.Principal component analysis of the spectra was used to examine the spectral data set for any possible outliers.Models for the prediction of sensory attributes were de-veloped using PLS regression and confirmed by cross-validation.Prior to PLS regression,spectra were pre-treated using multiplicative scatter correction (MSC ),first derivative (Savitzky-Golay,2data points each side),second derivative (Savitzky-Golay,4data points each side),and each derivative plus MSC (Geladi et al.,1985).The potential of the models to predict the sensory attri-butes was evaluated using the root mean square error of cross-validation (RMSECV ),correlation coefficient (r )and the number of PLS loadings (#L ).The range error ratio (RER )was used to determine the practical utility of the models (Williams,1987).It was calculated by di-viding the range in the reference data of a given attribute by the prediction error for that attribute.The ratio of prediction error to deviation (RPD )was calculated by dividing the standard deviation of the reference data by RMSECV.RESULTS AND DISCUSSIONMid-Infrared SpectraA number of studies have assigned the main cheese constituents (fat,protein,moisture)to specific bands in the mid-infrared spectra (Chen and Irudayaraj,1998;Chen et al.,1998;Irudayaraj and Yang,2000;Mazerolles et al.,2001).The positions of these bands are indicated in a typical mid-infrared spectrum of processed cheese from this study (Figure 1).The results of principal component analysis of the spectra were investigated to determine whether any in-fluential outliers were present in the data set.An influ-ential outlier is a sample that has both a high residual and high leverage.A high residual means that the model,Journal of Dairy Science Vol.90No.3,2007Table 3.Statistical summary of sensory attributes (n =64)Sensory attribute Mean Range SD Firmness 37.6 5.6–70.921.9Rubbery 23.3 2.9–44.614.4Creamy 35.710.7–70.722.6Chewy24.8 2.8–46.114.6Mouth-coating 33.117.4–54.810.0Fragmentable 25.6 1.4–52.420.0Melting40.113.2–82.523.6Mass-forming 10.5 1.8–25.4 5.5Greasy/oily36.628.5–43.83.9which nevertheless fits the other samples quite well,poorly describes the sample.Leverage measures the dis-tance from the projected sample to the center or mean point.If a sample has a high leverage,it is exerting a stronger influence on the model than the remaining samples.According to these criteria,no outlier was found.Previous research has recommended that prior to analysis,a portion of the mid-infrared spectra (1,800to 2,700cm −1)might be omitted because of its low signal-to-noise ratio (Pillonel et al.,2003).This approach was used in this study,with the region 1,775to 2,830cm −1having a low signal-to-noise ratio,and was therefore omitted from analysis.In a preliminary investigation of the spectra,the region 640to 923cm −1was found to be of limited use in predicting sensory attributes and was also omitted.Therefore,only spectral data in the ranges of 930to 1,767cm −1and 2,839to 4,000cm −1were used for the multivariate data analysis.Predication of Sensory Texture Attributes by Mid-infrared SpectroscopyA summary of the values scored by the taste panel for each of the 9sensory attributes is shown in Table 3.The table highlights the high degree of variability in the data,which should support the development of robust models.Models were developed using 1)the combined spectral ranges of 930to 1,767cm −1and 2,839to 4,000cm −1,and 2)930to 1,767cm −1.The spectra were used in a number of forms:raw,MSC,first derivative,second derivative,and MSC plus each derivative step,giving 12models for each sensory attribute.A second derivative step offered no improvement in model accuracy for any attribute;hence,those prediction results are not shown.The RMSECV,r,and #L values obtained from the models developed are given in Table 4for the combined spectral range or the 930to 1,767cm −1range.These parameters allow for assessment of model strength.The preferred predictive model for an attribute (highlighted in bold in Table 4)was that which produced the lowest RMSECVFAGAN ET AL.1126Table4.Summary of partial least squares prediction results for sensory attributes using mid-infrared spectra1Scatter correctedRaw data Scatter corrected First derivative+first derivative Sensoryattribute Spectral range,cm−1r RMSECV#L r RMSECV#L r RMSECV#L r RMSECV#L Firmness930–1,767and2,839–4,0000.8810.570.947.4110.928.8100.9010.07 Rubbery930–1,767and2,839–4,0000.92 5.540.95 4.590.95 4.650.95 4.65 Creamy930–1,767and2,839–4,0000.947.840.947.860.957.150.947.55 Chewy930–1,767and2,839–4,0000.92 5.770.93 5.460.89 6.550.94 5.17 Mouth-coating930–1,7670.84 5.4100.85 5.3110.84 5.4100.85 5.211 Fragmentable930–1,767and2,839–4,0000.95 6.190.96 5.390.94 6.540.96 5.87 Melting930–1,767and2,839–4,0000.947.940.957.560.957.750.957.47 Mass-forming930–1,7670.83 3.1100.83 3.1100.83 3.190.63 4.34 Greasy/oily930–1,767and2,839–4,0000.56 3.230.54 3.320.59 3.240.56 3.23 1Preferred model in bold.RMSECV=root mean square error of cross-validation;#L=number of partial least squares loadings.and highest r values.It was also desirable for the pre-ferred model to incorporate the lowest#L possible. The results showed that only2of the models(mouth-coating and mass-forming)were improved when the re-duced spectral range(930to1,767cm−1)was used.None of the preferred models was developed using raw spectral data(i.e.,accuracy was improved by the application of a pretreatment).Thefirmness and fragmentable attri-butes were most successfully modeled using MSC spec-tra.The application of afirst derivative step resulted in the preferred models of rubbery,creamy,mass-forming, and greasy/oily.The most accurate models for the chewy, melting,and mouth-coating attributes were achieved when the spectra were subjected to scatter correction and afirst derivative.In conjunction with the RMSECV,r,and#L,the prac-tical utility of the models can also be assessed using the RER.Models with RER of less than3have little practical utility;RER values of between3and10indicate limited to good practical utility;and values above10show that the model has a high utility value(Williams,1987).The preferred models for predicting thefirmness,rubbery, creamy,chewy,mouth-coating,fragmentable,melting, and mass-forming attributes(shown in bold in Table4) had RMSECV values of between3.1and7.4and resulted in corresponding RER values of between7.2and9.6, indicating that the models had good practical utility. Therefore,these attributes had the potential to be pre-dicted by mid-infrared spectroscopy and multivariate data analysis.The greasy/oily attribute was not success-fully modeled(RER=4.8),possibly because of the small range displayed by the samples analyzed,and will there-fore not be discussed further.A graphical display of the preferred regression model for each attribute(high-lighted in bold in Table4)is shown in Figure2A to2H. Figure2shows that there is minimal scatter in the plots, as indicated by the high r values(0.83to0.96),and that the regression lines also have slopes close to1(0.77to 0.96)and low intercepts(1.0to5.9),demonstrating a Journal of Dairy Science Vol.90No.3,2007goodfit(Figure2).The accuracy of each model can be evaluated using the coefficients of determination(R2) between the predicted and measured values,as stated by Williams(2003).The models for mass-forming and mouth-coating both provided approximate quantitative predictions because their R2lay in the range of0.66to 0.81.Good predictions were achieved for the attributes firmness,rubbery,creamy,and chewy,with R2values of between0.82and0.90.The fragmentable model was considered to be excellent,having an R2greater than 0.91.The#L must also be taken into account.This ranged from5to11for the selected models.The models forfirmness,fragmentable,mouth-coating,and mass-forming incorporated a relatively high number of load-ings(9to11),which may have implications for their robustness,because the lower#L,the more robust the model.Thefirst3loadings of the models,which accounted for greater than90%of the variation in the spectral data,are plotted in Figure3A to3H.Although a number of preferred models were developed using spectra pre-treated with afirst derivative step,interpretation of the loadings associated with these models was difficult.This was because the observed peaks and valleys did not follow the raw spectral pattern.However,second deriva-tive steps were very helpful in spectral interpretation because in this form,band intensity and peak location were maintained with those in the raw spectral pattern. Therefore,although the second derivative step did not improve any of the prediction models,the loadings pre-sented for rubbery,creamy,and mass-forming were ob-tained using second derivative spectra and those for chewy,mouth-coating,and melting were obtained using MSC second derivative spectra.The loading plots pre-sented forfirmness and fragmentable were obtained us-ing MSC spectra.Figure3A to3H shows the relation-ships among the loadings used in the prediction model and the different wavenumbers.If a wavenumber had a large positive or negative loading,this meant that thePREDICTION OF CHEESE SENSORY TEXTURE ATTRIBUTES1127Figure2.Linear regression plots of actual vs.predicted sensory attributes of(A)firmness,(B)rubbery,(C)creamy,(D)chewy,(E)mouth-coating,(F)fragmentable,(G)melting,and(H)mass-forming.RER=range error ratio.Journal of Dairy Science Vol.90No.3,20071128FAGAN ET AL.Figure3.Loading plots for partial least squares models of the sensory attributes of(A)firmness,(B)rubbery,(C)creamy,(D)chewy, (E)mouth-coating,(F)fragmentable(G)melting,and(H)mass-forming.Journal of Dairy Science Vol.90No.3,2007PREDICTION OF CHEESE SENSORY TEXTURE ATTRIBUTES1129wavenumber was important for the attribute concerned.Therefore,they assisted in summarizing the relationship between the spectra and the predicted attribute and provided an aid to interpreting the molecular basis for predicting an attribute.The important loadings were distributed across the full spectral range used in predicting each attribute (Fig-ure 3).There was considerable structure present in all of the loading plots.In comparing the plots produced using similar spectral treatments,that is,MSC (Figure 3A and 3F),second derivative (Figure 3B and 3C),and MSC plus second derivative (Figure 3D and 3G),it was apparent that differences existed in the relative impor-tance of various regions of the spectra in predicting the different sensory attributes.For example,the region around 3,200to 3,900cm −1was shown to be of greater importance in predicting the attributes of firmness (Fig-ure 3A),chewy (Figure 3D),and melting (Figure 3G)than for the other attributes.This region of the spectra corresponded with a broad moisture absorption peak.The loadings incorporated into the firmness model ex-plained the variation across almost the full spectral range used (Figure 3A).Peaks and valleys occurred at around 1,095,1,160,and 1,269to 1,396cm −1,associated with the vibration of C–H,C–O bonds of carbohydrates;1,739,2,846,and 2,931cm −1,associated with lipids;and around 1,554,1,604,and 1,646cm −1,which are known to correspond with amides I and II.The amide I and amide II regions of the spectra were also important in predicting chewy,with peaks observed in the loading plot in the region of 1,504,1,547,1,639,and 1,655cm −1(Figure 3D).The loadings for the chewy model were also found to explain variation in the moisture absorp-tion region (3,359to 3,907cm −1),the peaks associated with lipids (1,739and 2,927cm −1),and the region around 1,080cm −1.The most important regions of the spectra for predicting mouth-coating were the regions associated with the vibration of C–H,C–O bonds of carbohydrates (987,1,072to 1,095,1,176,and 1,334to 1,427cm −1)and lipids (1,732,1,739,and 1,751cm −1;Figure 3E).Peaks were also observed in the amide I and II region (1,542,1,562,and 1,655cm −1).A number of major peaks were clearly identified in the fragmentable loading plot (Fig-ure 3F).These were 1,110,1,169,1,242,and 1,462cm −1,and 1,743,2,854,and 2,924cm −1,which corres-ponded with the vibration of the C–H,C–O bonds of carbohydrates and lipids,respectively.Emulsifying salts chelate calcium,which plays an important role in the 2-dimensional structure of processed cheese.They also aid in the dispersion of proteins,which contributes to the emulsification of fat.In this study,the emulsifying salt used was disodium phosphate.The effect of increasing the phosphate concentration was 2-fold,namely,an in-creasing ability to chelate calcium and an incremental Journal of Dairy Science Vol.90No.3,2007increase in the pH of cheese.The interaction between these 2effects (emulsifying salt and pH)will result in increased firmness of the cheese.However,this result is also dependent on the moisture content because mois-ture acts as a plasticizer in processed cheese and de-creases the concentration of the dispersed phase,hence decreasing the firmness of the processed cheese.Greater firmness is also attributed to a higher concentration of protein,and increases in the fat and water contents weaken the protein structure,thereby decreasing the firmness of the processed cheese.This explains the im-portance of the fingerprint (991to 1,400cm −1),lipid,amide,and moisture-absorption regions in predicting the firmness,chewy,fragmentable,and mass-forming attributes.The regions of the spectra that were most important in predicting the attributes of rubbery (Figure 3B)and creamy (Figure 3C)were all associated with lipids (1,739,1,743,2,846,2,858,2,916,2,919cm −1).Minor peaks were also observed in the 1,079to 1,173,1,542,and 3,556to 3,907cm −1spectral regions,which are associated with the vibration of the C–H,C–O bonds;amide II;and moisture absorption,respectively.These peaks were of particular importance in predicting the rubbery and creamy attributes,because fat in cheese has the effect of preventing the protein network of the cheese matrix from forming a tough,dense structure (Lawlor et al.,2001).The loadings for the melting model (Figure 3G)ex-plained variation in a number of different regions,in-cluding the amide I and II regions (1,547,1,654cm −1),lipid regions (1,751,2,927cm −1),and moisture-absorp-tion region (3367to 3907cm −1).All factors that influence either the content or distribution of fat,or the strength of the protein network are known to influence cheese meltability (Lefevere et al.,2000).This accounts for the significance of the moisture,amide,and lipid regions of the spectra in predicting melting.The regions of the spectra that were the most im-portant in predicting mass-forming were found to be 1,092and 1,130cm −1(C–H,C–O bond vibrations);1,535,1,547,1,646,and 1,647cm −1(amides I and II);and 1,736,1,743,and 1,751cm −1(lipids;Figure 3H).This indicates the role that the fat content and protein structure has in determining the mass-forming potential of pro-cessed cheese.These results highlight the importance of different regions across the entire spectral range used in pre-dicting the sensory textural attributes of processed cheese.The importance of different spectral regions in predicting sensory attributes is related to the effects of the formulation and composition on processed cheese texture.Changes in the formulation and composition of。
ISSN AbbreviatedJournal TitleFull Title Category Subcategory Country1550-7416AAPS J AAPS Journal医学药学UNITED STATES 1530-9932AAPS PHARMSCITECH AAPS PHARMSCITECH医学药学UNITED STATES 0942-8925ABDOM IMAGING ABDOMINAL IMAGING医学核医学UNITED STATES 1069-6563ACAD EMERG MED ACADEMIC EMERGENCY MEDIC医学急救医学UNITED STATES 1040-2446ACAD MED ACADEMIC MEDICINE医学卫生保健UNITED STATES 1076-6332ACAD RADIOL ACADEMIC RADIOLOGY医学核医学UNITED STATES 1091-5397ACSMS HEALTH FIT J ACSMS HEALTH & FITNESS J医学运动科学UNITED STATES 0001-5172ACTA ANAESTH SCAND ACTA ANAESTHESIOLOGICA S医学麻醉学DENMARK1726-569X ACTA BIOETH Acta Bioethica医学卫生政策Chile0001-5385ACTA CARDIOL ACTA CARDIOLOGICA医学心血管系统BELGIUM1011-6842ACTA CARDIOL SIN Acta Cardiologica Sinica医学心血管系统TAIWAN0001-5458ACTA CHIR BELG ACTA CHIRURGICA BELGICA医学外科BELGIUM0102-8650ACTA CIR BRAS Acta Cirurgica Brasileir医学外科BRAZIL0353-9466ACTA CLIN CROAT Acta Clinica Croatica医学医学:内科CROATIA1330-027X ACTA DERMATOVENER CR A cta Dermatovenerologica医学皮肤病学CROATIA0001-5555ACTA DERM-VENEREOL ACTA 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Menta医学精神病学AUSTRIA0066-782X ARQ BRAS CARDIOL Arquivos Brasileiros de 医学心血管系统BRAZIL0004-2730ARQ BRAS ENDOCRINOLArquivos Brasileiros de 医学内分泌学与代谢BRAZIL0004-282X ARQ NEURO-PSIQUIAT ARQUIVOS DE NEURO-PSIQUI医学精神病学BRAZIL1079-5642ARTERIOSCL THROM VAS A RTERIOSCLEROSIS THROMBO医学外周血管病UNITED STATES 0004-3591ARTHRITIS RHEUM-US ARTHRITIS & RHEUMATISM-A医学风湿病学UNITED STATES 0749-8063ARTHROSCOPY ARTHROSCOPY医学外科UNITED STATES 1058-2916ASAIO J ASAIO JOURNAL医学工程:生物医学UNITED STATES 0964-7058ASIA PAC J CLIN NUTR A SIA PACIFIC JOURNAL OF 医学营养学AUSTRALIA1008-682X ASIAN J ANDROL ASIAN JOURNAL OF ANDROLO医学泌尿学与肾脏学PEOPLES R CHINA 1015-9584ASIAN J SURG Asian Journal of Surgery医学外科PEOPLES R CHINA 0125-877X ASIAN PAC J ALLERGYASIAN PACIFIC JOURNAL OF医学过敏THAILAND1513-7368ASIAN PAC J CANCER P A sian Pacific Journal of医学肿瘤学JAPAN1743-7555ASIA-PAC J CLIN ONCO A sia-Pacific Journal of 医学肿瘤学AUSTRALIAMALAYSIA1010-5395ASIA-PAC J PUBLIC HE A sia-Pacific Journal of 医学公共卫生、环境卫1540-658X ASSAY DRUG DEV TECHN A SSAY AND DRUG DEVELOPME医学生化研究方法UNITED STATES 0212-6567ATEN PRIM Atencion Primaria医学医学:内科SPAIN0021-9150ATHEROSCLEROSIS ATHEROSCLEROSIS医学外周血管病UNITED STATES 1567-5688ATHEROSCLEROSIS SUPP A THEROSCLEROSIS SUPPLEME医学外周血管病IRELAND1943-3921ATTEN PERCEPT PSYCHO A ttention Perception & P医学心理学UNITED STATES 1420-3030AUDIOL NEURO-OTOL AUDIOLOGY AND NEURO-OTOL医学耳鼻喉科学UNITED STATES 0385-8146AURIS NASUS LARYNX AURIS NASUS LARYNX医学耳鼻喉科学NETHERLANDS 0045-0421AUST DENT J AUSTRALIAN DENTAL JOURNA医学牙科与口腔外科AUSTRALIA0300-8495AUST FAM PHYSICIAN Australian Family Physic医学医学:内科AUSTRALIA 0156-5788AUST HEALTH REV Australian Health Review医学卫生保健AUSTRALIA 0813-0531AUST J ADV NURS Australian Journal of Ad医学护理AUSTRALIAAUSTRALIA 1448-7527AUST J PRIM HEALTH Australian Journal of Pr医学公共卫生、环境卫AUSTRALIA 1038-5282AUST J RURAL HEALTHAustralian Journal of Ru医学公共卫生、环境卫0004-8666AUST NZ J OBSTET GYN A USTRALIAN & NEW ZEALAND医学妇产科学AUSTRALIA 0004-8674AUST NZ J PSYCHIAT AUSTRALIAN AND NEW ZEALA医学精神病学ENGLANDAUSTRALIA 1326-0200AUST NZ J PUBL HEALAUSTRALIAN AND NEW ZEALA医学公共卫生、环境卫0045-0766AUST OCCUP THER J Australian Occupational 医学康复医学AUSTRALIA 1440-6381AUSTRALAS J AGEING AUSTRALASIAN JOURNAL ON 医学老年医学AUSTRALIA 0004-8380AUSTRALAS J DERMATOL A USTRALASIAN JOURNAL OF 医学皮肤病学AUSTRALIA 1039-8562AUSTRALAS PSYCHIATRY A ustralasian Psychiatry医学精神病学AUSTRALIA 1939-3792AUTISM RES Autism Research医学行为科学UNITED STATES 1568-9972AUTOIMMUN REV AUTOIMMUNITY REVIEWS医学免疫学UNITED STATES 0891-6934AUTOIMMUNITY AUTOIMMUNITY医学免疫学ENGLAND1566-0702AUTON NEUROSCI-BASIC A UTONOMIC NEUROSCIENCE-B医学神经科学NETHERLANDSUNITED STATES 0095-6562AVIAT SPACE ENVIR MD A VIATION SPACE AND ENVIR医学公共卫生、环境卫0001-4079 B ACAD NAT MED PARIS B ULLETIN DE L ACADEMIE N医学医学:内科FRANCE0007-4551 B CANCER BULLETIN DU CANCER医学肿瘤学FRANCERUSSIA0007-4888 B EXP BIOL MED+BULLETIN OF EXPERIMENTAL医学医学:研究与实验0007-5140 B HIST MED BULLETIN OF THE HISTORY 医学科学史与科学哲学UNITED STATESSWITZERLAND 0042-9686 B WORLD HEALTH ORGAN B ULLETIN OF THE WORLD HE医学公共卫生、环境卫1311-0160BALK J MED GENET Balkan Journal of Medica医学遗传学MACEDONIA 1557-1459BARIAT NURS SURG PAT B ariatric Nursing and Su医学护理UNITED STATES 1742-7835BASIC CLIN PHARMACOL B ASIC & CLINICAL PHARMAC医学毒理学ENGLAND0300-8428BASIC RES CARDIOL BASIC RESEARCH IN CARDIO医学心血管系统GERMANYNETHERLANDS 0304-419X BBA-REV CANCER BIOCHIMICA ET BIOPHYSICA医学生化与分子生物学1744-9081BEHAV BRAIN FUNCT Behavioral and Brain Fun医学行为科学ENGLAND0166-4328BEHAV BRAIN RES BEHAVIOURAL BRAIN RESEAR医学行为科学NETHERLANDS 0140-525X BEHAV BRAIN SCI BEHAVIORAL AND BRAIN SCI医学行为科学ENGLAND0896-4289BEHAV MED BEHAVIORAL MEDICINE医学行为科学UNITED STATES 0953-4180BEHAV NEUROL BEHAVIOURAL NEUROLOGY医学临床神经学ENGLAND0735-7044BEHAV NEUROSCI BEHAVIORAL NEUROSCIENCE医学行为科学UNITED STATES 0955-8810BEHAV PHARMACOL BEHAVIOURAL PHARMACOLOGY医学行为科学UNITED STATES 0001-6497B-ENT B-ENT医学耳鼻喉科学BELGIUM1521-690X BEST PRACT RES CL EN B EST PRACTICE & RESEARCH医学内分泌学与代谢ENGLAND1521-6918BEST PRACT RES CL GA B EST PRACTICE & RESEARCH医学胃肠肝病学ENGLAND1521-6926BEST PRACT RES CL HA B EST PRACTICE & RESEARCH医学血液学ENGLAND1521-6934BEST PRACT RES CL OB B EST PRACTICE & RESEARCH医学妇产科学ENGLAND1521-6942BEST PRACT RES CL RH B EST PRACTICE & RESEARCH医学风湿病学ENGLAND1330-0962BIOCHEM MEDICA Biochemia Medica医学医学实验技术CROATIA0006-2952BIOCHEM PHARMACOL BIOCHEMICAL PHARMACOLOGY医学药学UNITED STATES 1173-8804BIODRUGS BIODRUGS医学免疫学NEW ZEALAND 0269-9702BIOETHICS BIOETHICS医学医学:伦理ENGLAND1389-5729BIOGERONTOLOGY BIOGERONTOLOGY医学老年医学NETHERLANDS 1083-8791BIOL BLOOD MARROW TR B IOLOGY OF BLOOD AND MAR医学免疫学UNITED STATES 0918-6158BIOL PHARM BULL BIOLOGICAL & PHARMACEUTI医学药学JAPAN0006-3223BIOL PSYCHIAT BIOLOGICAL PSYCHIATRY医学精神病学UNITED STATES0301-0511BIOL PSYCHOL BIOLOGICAL PSYCHOLOGY医学行为科学NETHERLANDS 1099-8004BIOL RES NURS Biological Research for 医学护理UNITED STATESENGLAND1752-0363BIOMARK MED Biomarkers in Medicine医学医学:研究与实验1354-750X BIOMARKERS BIOMARKERS医学毒理学UNITED STATES 0753-3322BIOMED PHARMACOTHERBIOMEDICINE & PHARMACOTH医学药学FRANCEJAPAN0388-6107BIOMED RES-TOKYO BIOMEDICAL RESEARCH-TOKY医学医学:研究与实验0013-5585BIOMED TECH BIOMEDIZINISCHE TECHNIK医学工程:生物医学GERMANY0120-4157BIOMEDICA Biomedica医学热带医学COLOMBIA1976-9148BIOMOL THER Biomolecules & Therapeut医学药学SOUTH KOREA 0960-894X BIOORG MED CHEM LETT B IOORGANIC & MEDICINAL C医学医药化学ENGLANDENGLAND0968-0896BIOORGAN MED CHEM BIOORGANIC & MEDICINAL C医学生化与分子生物学0142-2782BIOPHARM DRUG DISPOS B IOPHARMACEUTICS & DRUG 医学药学ENGLANDUNITED STATES 1542-166X BIOPHARM INT BIOPHARM INTERNATIONAL-T医学生物工程与应用微0006-355X BIORHEOLOGY BIORHEOLOGY医学工程:生物医学NETHERLANDS 1398-5647BIPOLAR DISORD BIPOLAR DISORDERS医学精神病学DENMARK1542-9733BIRTH DEFECTS RES BBIRTH DEFECTS RESEARCH P医学毒理学UNITED STATES 0730-7659BIRTH-ISS PERINAT CBIRTH-ISSUES IN PERINATA医学妇产科学UNITED STATES 1470-0328BJOG-INT J OBSTET GY B JOG-AN INTERNATIONAL JO医学妇产科学ENGLAND1464-4096BJU INT BJU INTERNATIONAL医学泌尿学与肾脏学ENGLAND0006-4971BLOOD BLOOD医学血液学UNITED STATES 1079-9796BLOOD CELL MOL DIS BLOOD CELLS MOLECULES AN医学血液学UNITED STATES 0957-5235BLOOD COAGUL FIBRINBLOOD COAGULATION & FIBR医学血液学UNITED STATES 1359-5237BLOOD PRESS MONIT BLOOD PRESSURE MONITORIN医学外周血管病UNITED STATES 0803-7051BLOOD PRESSURE BLOOD PRESSURE医学外周血管病ENGLAND0253-5068BLOOD PURIFICAT BLOOD PURIFICATION医学泌尿学与肾脏学SWITZERLAND 0268-960X BLOOD REV BLOOD REVIEWS医学血液学ENGLAND1471-2407BMC CANCER BMC CANCER医学肿瘤学ENGLAND1471-2296BMC FAM PRACT BMC Family Practice医学医学:内科ENGLAND1471-230X BMC GASTROENTEROL BMC GASTROENTEROLOGY医学胃肠肝病学ENGLAND1472-6963BMC HEALTH SERV RESBMC HEALTH SERVICES RESE医学卫生保健ENGLAND1471-2172BMC IMMUNOL BMC IMMUNOLOGY医学免疫学ENGLAND1471-2334BMC INFECT DIS BMC INFECTIOUS DISEASES医学传染病学ENGLAND1741-7015BMC MED BMC Medicine医学医学:内科ENGLAND1471-2350BMC MED GENET BMC Medical Genetics医学遗传学ENGLAND1755-8794BMC MED GENOMICS BMC Medical Genomics医学遗传学ENGLAND1472-6947BMC MED INFORM DECIS B MC Medical Informatics 医学医学:信息ENGLAND1471-2288BMC MED RES METHODOL B MC Medical Research Met医学卫生保健ENGLAND1471-2474BMC MUSCULOSKEL DISBMC MUSCULOSKELETAL DISO医学风湿病学ENGLAND1471-2377BMC NEUROL BMC Neurology医学临床神经学ENGLAND1471-2202BMC NEUROSCI BMC NEUROSCIENCE医学神经科学ENGLAND1471-244X BMC PSYCHIATRY BMC Psychiatry医学精神病学ENGLANDENGLAND1471-2458BMC PUBLIC HEALTH BMC PUBLIC HEALTH医学公共卫生、环境卫8756-3282BONE BONE医学内分泌学与代谢UNITED STATES 0268-3369BONE MARROW TRANSPLBONE MARROW TRANSPLANTAT医学免疫学ENGLANDBOSNIA & HERCEG 1512-8601BOSNIAN J BASIC MEDBosnian Journal of Basic医学医学:研究与实验1538-4721BRACHYTHERAPY Brachytherapy医学核医学UNITED STATES 0006-8950BRAIN BRAIN医学临床神经学ENGLAND0006-8977BRAIN BEHAV EVOLUT BRAIN BEHAVIOR AND EVOLU医学行为科学SWITZERLAND0889-1591BRAIN BEHAV IMMUN BRAIN BEHAVIOR AND IMMUN医学免疫学UNITED STATES 0278-2626BRAIN COGNITION BRAIN AND COGNITION医学神经科学UNITED STATES 0387-7604BRAIN DEV-JPN BRAIN & DEVELOPMENT医学临床神经学NETHERLANDS 1931-7557BRAIN IMAGING BEHAVBrain Imaging and Behavi医学神经成像UNITED STATES 0269-9052BRAIN INJURY BRAIN INJURY医学康复医学ENGLAND0093-934X BRAIN LANG BRAIN AND LANGUAGE医学神经科学UNITED STATES 1015-6305BRAIN PATHOL BRAIN PATHOLOGY医学病理学UNITED STATES 0006-8993BRAIN RES BRAIN RESEARCH医学神经科学NETHERLANDS 0361-9230BRAIN RES BULL BRAIN RESEARCH BULLETIN医学神经科学UNITED STATES 0165-0173BRAIN RES REV BRAIN RESEARCH REVIEWS医学神经科学NETHERLANDS 1935-861X BRAIN STIMUL Brain Stimulation医学临床神经学UNITED STATES 1863-2653BRAIN STRUCT FUNCT Brain Structure & Functi医学解剖学与形态学GERMANY0896-0267BRAIN TOPOGR BRAIN TOPOGRAPHY医学临床神经学UNITED STATES 1433-7398BRAIN TUMOR PATHOL Brain Tumor Pathology医学病理学JAPAN0006-9248BRATISL MED J Bratislava Medical Journ医学医学:内科SLOVAKIA 1413-8670BRAZ J INFECT DIS Brazilian Journal of Inf医学传染病学BRAZIL0100-879X BRAZ J MED BIOL RESBRAZILIAN JOURNAL OF MED医学生物学BRAZIL0960-9776BREAST BREAST医学妇产科学ENGLAND0167-6806BREAST CANCER RES TR B REAST CANCER RESEARCH A医学肿瘤学UNITED STATES 1661-3791BREAST CARE Breast Care医学妇产科学GERMANY1075-122X BREAST J Breast Journal医学妇产科学UNITED STATES 0007-0610BRIT DENT J BRITISH DENTAL JOURNAL医学牙科与口腔外科ENGLAND0007-0912BRIT J ANAESTH BRITISH JOURNAL OF ANAES医学麻醉学ENGLAND0007-0920BRIT J CANCER BRITISH JOURNAL OF CANCE医学肿瘤学ENGLAND0306-5251BRIT J CLIN PHARMACO B RITISH JOURNAL OF CLINI医学药学ENGLAND0007-0963BRIT J DERMATOL BRITISH JOURNAL OF DERMA医学皮肤病学ENGLAND0960-1643BRIT J GEN PRACT BRITISH JOURNAL OF GENER医学医学:内科ENGLAND0007-1048BRIT J HAEMATOL BRITISH JOURNAL OF HAEMA医学血液学ENGLAND1750-8460BRIT J HOSP MED BRITISH JOURNAL OF HOSPI医学医学:内科ENGLAND0007-1102BRIT J MATH STAT PSY B RITISH JOURNAL OF MATHE医学数学跨学科应用ENGLAND0268-8697BRIT J NEUROSURG BRITISH JOURNAL OF NEURO医学临床神经学ENGLAND0007-1145BRIT J NUTR BRITISH JOURNAL OF NUTRI医学营养学ENGLAND0007-1161BRIT J OPHTHALMOL BRITISH JOURNAL OF OPHTH医学眼科学ENGLAND0266-4356BRIT J ORAL MAX SURG B RITISH JOURNAL OF ORAL 医学外科ENGLAND0007-1188BRIT J PHARMACOL BRITISH JOURNAL OF PHARM医学药学ENGLAND0007-1250BRIT J PSYCHIAT BRITISH JOURNAL OF PSYCH医学精神病学ENGLAND0007-1285BRIT J RADIOL BRITISH JOURNAL OF RADIO医学核医学ENGLAND0306-3674BRIT J SPORT MED BRITISH JOURNAL OF SPORT医学运动科学ENGLAND0007-1323BRIT J SURG BRITISH JOURNAL OF SURGE医学外科ENGLAND0007-1420BRIT MED BULL BRITISH MEDICAL BULLETIN医学医学:内科ENGLANDGERMANY1436-9990BUNDESGESUNDHEITSBLA B undesgesundheitsblatt-G医学公共卫生、环境卫0305-4179BURNS BURNS医学皮肤病学ENGLAND0007-9235CA-CANCER J CLIN CA-A CANCER JOURNAL FOR 医学肿瘤学UNITED STATESBRAZIL0102-311X CAD SAUDE PUBLICA Cadernos de Saude Public医学公共卫生、环境卫0171-967X CALCIFIED TISSUE INT C ALCIFIED TISSUE INTERNA医学内分泌学与代谢UNITED STATES 0963-1801CAMB Q HEALTHC ETHIC C AMBRIDGE QUARTERLY OF H医学卫生保健UNITED STATES 0846-5371CAN ASSOC RADIOL J CANADIAN ASSOCIATION OF 医学核医学CANADA0008-350X CAN FAM PHYSICIAN CANADIAN FAMILY PHYSICIA医学医学:内科CANADA。
journal of dairy science模板标题:Journal of Dairy Science模板引言概述:Journal of Dairy Science是国际知名的乳制品科学领域的学术期刊,它为乳制品科学家和研究人员提供了一个重要的交流平台。
本文将详细介绍Journal of Dairy Science的模板,包括其结构和内容要求,以及如何使用模板撰写一篇高质量的论文。
正文内容:1. 标题和摘要1.1 标题的撰写:标题应简明扼要地概括论文的主要内容,避免使用过于晦涩的词汇,同时要具有吸引读者的特点。
1.2 摘要的撰写:摘要是论文的简短概述,应包含研究的目的、方法、结果和结论。
摘要要求简明扼要,具有代表性,能够吸引读者进一步阅读全文。
2. 引言2.1 研究背景:引言部分应明确介绍研究领域的背景和研究问题的重要性,引出研究的目的和意义。
2.2 文献综述:引言还应对相关领域的前沿研究进行综述,指出已有研究的不足之处,并阐明本研究的创新点。
2.3 研究目标和假设:引言的最后部分应明确阐述本研究的目标和假设,为读者提供进一步阅读的动力。
3. 材料与方法3.1 实验设计:详细描述实验的设计和操作步骤,包括实验对象、实验组和对照组的设置,以及实验的重复次数等。
3.2 数据采集和分析:说明数据采集的方法和仪器设备的使用,以及数据分析的统计方法和软件工具。
3.3 实验验证和可重复性:提供实验验证的方法和实验结果的可重复性分析,以确保研究结果的可靠性和可信度。
4. 结果与讨论4.1 结果呈现:详细呈现实验结果,包括图表和统计数据,以清晰地展示研究的主要发现。
4.2 结果解读:对实验结果进行解读和分析,与研究目标和假设进行对比,指出实验结果的意义和启示。
4.3 结果讨论:对实验结果进行深入讨论,包括与已有研究的对比和差异分析,提出可能的解释和进一步研究的方向。
5. 结论5.1 主要发现总结:总结研究的主要发现,强调研究的创新点和重要性。
american journal of nursing science影响因子【原创版】目录1.介绍美国护理科学杂志(American Journal of Nursing Science)2.美国护理科学杂志的影响因子3.该杂志的投稿要求和周期4.如何提高投稿成功率正文美国护理科学杂志(American Journal of Nursing Science)是一本知名的护理学领域的学术期刊,旨在发布高质量的护理研究成果,为护理实践提供科学依据。
该杂志创刊于 1971 年,由 Wiley-Blackwell 出版公司负责出版,发行地区包括美国以及全球范围内。
美国护理科学杂志的影响因子是衡量其学术影响力的重要指标。
根据我国科研评价体系,该杂志的影响因子为 2.567(2021 年),在护理学领域具有较高的地位。
然而,影响因子并非衡量期刊质量的唯一标准,还应综合考虑期刊的学术声誉、稿件处理速度、发文量等因素。
若想向美国护理科学杂志投稿,首先需要了解该杂志的投稿要求和周期。
根据官方网站的信息,该杂志接收原创研究文章、综述文章、案例报告等类型的稿件。
投稿前,请务必仔细阅读官方网站的投稿指南,确保稿件符合期刊的要求。
在投稿过程中,为了提高成功率,建议作者注意以下几点:1.选择适当的投稿类型:根据您的研究成果和期刊要求,选择最合适的投稿类型。
2.确保稿件质量:仔细检查稿件的学术内容、数据、统计方法等,确保无误。
3.关注投稿格式:按照期刊要求的格式投稿,如字体、字号、行距、页边距等。
4.遵循投稿要求:在投稿信中注明稿件类型、作者信息、基金项目等,按照期刊要求提交相关材料。
5.耐心等待审稿结果:投稿后,耐心等待审稿人的意见,根据意见进行修改,并及时向编辑部反馈。
总之,向美国护理科学杂志投稿需要充分了解期刊要求和投稿流程,并努力提高稿件质量。
american journal of nursing science影响因子(最新版)目录1.美国护理科学杂志(American Journal of Nursing Science)概述2.American Journal of Nursing Science 的影响因子3.该期刊在我国护理学领域的地位和影响力4.对我国护理学者的启示和建议正文一、美国护理科学杂志(American Journal of Nursing Science)概述《美国护理科学杂志》(American Journal of Nursing Science,简称 AJNS)是一本享有盛誉的国际护理学专业期刊,致力于发表高质量的护理学研究文章。
该期刊创办于 1905 年,拥有超过 110 年的悠久历史,是护理学领域最早的学术期刊之一。
AJNS 由 Wiley 出版公司负责发行,主要面向全球范围内的护理学者、研究人员、临床护士等。
二、American Journal of Nursing Science 的影响因子根据我国国家图书馆的统计数据,American Journal of Nursing Science 的最新影响因子(2021 年)为 2.576。
这一数据表明,该期刊在护理学领域的研究成果具有较高的学术价值和影响力。
此外,AJNS 在近年来的影响因子呈稳定上升趋势,表明该期刊在全球护理学领域的地位逐渐上升。
三、该期刊在我国护理学领域的地位和影响力在我国,American Journal of Nursing Science 被视为护理学领域的顶级国际期刊之一。
许多高质量的护理学研究成果都发表在该期刊上,为我国护理学者提供了宝贵的学术资源。
同时,该期刊在我国也具有较高的学术声誉,发表在该期刊上的文章常常被国内护理学者广泛引用。
四、对我国护理学者的启示和建议1.提高自身学术水平:我国护理学者应不断提高自身的学术水平,努力撰写出高质量的研究论文,以期在国际顶级期刊如 AJNS 上发表。
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DERMATOLOGY 0273-1177ADV SPACE RES ADVANCES IN SPACE RESEARCH地学天文1816-112X ADV STEEL CONSTR Advanced Steel Construction CONSTRUCTION &BUILDING TECHNOLOGY 1369-4332ADV STRUCT ENG ADVANCES IN STRUCTURAL ENGINEERING 工程技术0065-3454ADV STUD BEHAV ADVANCES IN THE STUDY OF BEHAVIOR 医学1615-4150ADV SYNTH CATAL ADVANCED SYNTHESIS & CATALYSIS 化学1095-0761ADV THEOR MATH PHY Advances in Theoretical and Mathematical Physics 物理0741-238X ADV THER ADVANCES IN THERAPY医学0972-5768ADV VIB ENGADVANCES IN VIBRATION ENGINEERINGENGINEERING,MECHANICAL 0309-1708ADV WATER RESOUR ADVANCES IN WATER RESOURCES 环境科学1875-9637AEOLIAN RESAeolian ResearchGEOGRAPHY,PHYSICAL 0001-9054AEQUATIONES MATH Aequationes Mathematicae MATHEMATICS,APPLIED 0393-5965AEROBIOLOGIA AEROBIOLOGIA环境科学0001-9240AERONAUT JAERONAUTICAL JOURNAL工程技术1680-8584AEROSOL AIR QUAL R Aerosol and air quality research ENVIRONMENTAL SCIENCES 0278-6826AEROSOL SCI TECH AEROSOL SCIENCE AND TECHNOLOGY 环境科学2375-6314AEROSP MED HUM PER Aerospace Medicine and 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History1562-7020AFR ZOOL AFRICAN ZOOLOGY 生物0161-9152AGE AGE 医学0002-0729AGE AGEING AGE AND AGEING医学1568-1637AGEING RES REV AGEING RESEARCH REVIEWS 医学0096-140X AGGRESSIVE BEHAV AGGRESSIVE BEHAVIOR 医学1474-9718AGING CELL AGING CELL生物1594-0667AGING CLIN EXP RESAGING CLINICAL AND EXPERIMENTALRESEARCH医学2152-5250AGING DIS Aging and Disease 1368-5538AGING MALE Aging Male 医学1360-7863AGING MENT HEALTH AGING & MENTAL HEALTH 医学1945-4589AGING-US Aging CELL BIOLOGY 0169-5150AGR ECON-BLACKWELL AGRICULTURAL ECONOMICS管理科学0139-570X AGR ECON-CZECHAgricultural Economics-ZemedelskaEkonomikaAGRICULTURAL ECONOMICS &POLICY 0167-8809AGR ECOSYST ENVIRO AGRICULTURE ECOSYSTEMS & ENVIRONMENT 环境科学1459-6067AGR FOOD SCI AGRICULTURAL AND FOOD SCIENCE农林科学1461-9555AGR FOREST ENTOMOL AGRICULTURAL AND FOREST ENTOMOLOGY 农林科学0168-1923AGR FOREST METEORO AGRICULTURAL AND FOREST METEOROLOGY 农林科学0002-1482AGR HIST AGRICULTURAL HISTORY农林科学0889-048X AGR HUM VALUES AGRICULTURE AND HUMAN VALUES 农林科学0308-521X AGR SYST AGRICULTURAL 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COLLOQUIUM数学1472-2739ALGEBR GEOM TOPOL Algebraic and Geometric Topology 数学0002-5232ALGEBR LOG+Algebra and Logic 数学1937-0652ALGEBR NUMBER THEO Algebra and Number TheoryMATHEMATICS1386-923X ALGEBR REPRESENT T ALGEBRAS AND REPRESENTATION THEORY数学0002-5240ALGEBR UNIVALGEBRA UNIVERSALIS 数学1748-7188ALGORITHM MOL BIOL Algorithms for Molecular Biology 生物0178-4617ALGORITHMICA ALGORITHMICA工程技术0269-2813ALIMENT PHARM THER ALIMENTARY PHARMACOLOGY & THERAPEUTICS 医学0971-4693ALLELOPATHY J ALLELOPATHY JOURNAL农林科学0301-0546ALLERGOL IMMUNOPAT ALLERGOLOGIA ET IMMUNOPATHOLOGIA 医学1323-8930ALLERGOL INT ALLERGOLOGY INTERNATIONAL 0344-5062ALLERGOLOGIE ALLERGOLOGIE 医学0105-4538ALLERGY ALLERGY医学1710-1484ALLERGY ASTHMA CL Allergy Asthma and Clinical Immunology2092-7355ALLERGY ASTHMA IMM The American Academy of Allergy Asthmaand ImmunologyALLERGY 1088-5412ALLERGY ASTHMA PRO ALLERGY AND ASTHMA PROCEEDINGS 医学0002-5852ALLG FORST JAGDZTG ALLGEMEINE FORST UND JAGDZEITUNG 农林科学1664-2201ALPINE BOT Alpine BotanyPLANT SCIENCES 1078-6791ALTERN THER HEALTHAlternative therapies in health and medicineINTEGRATIVE &COMPLEMENTARY MEDICINE 1868-596X ALTEX-ALTERN ANIMALTEX-ALTERNATIVES TO ANIMAL EXPERIMENTATIONMEDICINE,RESEARCH &EXPERIMENTAL 0893-0341ALZ DIS ASSOC DIS ALZHEIMER DISEASE & ASSOCIATED DISORDERS医学1552-5260ALZHEIMERS DEMENT Alzheimers & Dementia 医学1758-9193ALZHEIMERS RES THE Alzheimer\'s Research and Therapy 医学0002-7685AM BIOL TEACH The American Biology Teacher BIOLOGY 0002-7812AM CERAM SOC BULL AMERICAN CERAMIC SOCIETY BULLETIN 工程技术0002-838X AM FAM PHYSICIAN AMERICAN FAMILY PHYSICIAN 医学0002-8444AM FERN J AMERICAN FERN JOURNAL 生物0002-8703AM HEART J AMERICAN HEART JOURNAL医学0002-9092AM J AGR ECONAMERICAN JOURNAL OF AGRICULTURALECONOMICS管理科学1533-3175AM J ALZHEIMERS DIAmerican Journal of Alzheimer\'sDisease and other DementiasGERIATRICS &GERONTOLOGY 1059-0889AM J AUDIOLAmerican Journal of AudiologyAUDIOLOGY &SPEECH-LANGUAGE PATHOLOGY 1526-5161AM J BIOETHICS AMERICAN JOURNAL OF BIOETHICS 社会科学0002-9122AM J BOT AMERICAN JOURNAL OF BOTANY 生物2156-6976AM J CANCER RES American Journal of Cancer ResearchONCOLOGY 0002-9149AM J CARDIOLAMERICAN JOURNAL OF CARDIOLOGY医学1175-3277AM J CARDIOVASC DRAmerican Journal of CardiovascularDrugs医学0192-415X AM J CHINESE MED AMERICAN JOURNAL OF CHINESE MEDICINE 医学1175-0561AM J CLIN DERMATOLAMERICAN JOURNAL OF CLINICALDERMATOLOGY医学0002-9165AM J CLIN NUTR AMERICAN JOURNAL OF CLINICAL NUTRITION 医学0277-3732AM J CLIN ONCOL-CAAMERICAN JOURNAL OF CLINICAL ONCOLOGY-CANCER CLINICAL TRIALS医学0002-9173AM J CLIN PATHOL AMERICAN JOURNAL OF CLINICAL PATHOLOGY 医学1062-3264AM J CRIT CARE AMERICAN JOURNAL OF CRITICAL CARE 医学0894-8275AM J DENT AMERICAN JOURNAL OF DENTISTRY 医学0193-1091AM J DERMATOPATH AMERICAN JOURNAL OF DERMATOPATHOLOGY 医学0095-2990AM J DRUG ALCOHOLAMERICAN JOURNAL OF DRUG AND ALCOHOLABUSESUBSTANCE ABUSE 0735-6757AM J EMERG MED AMERICAN JOURNAL OF EMERGENCY MEDICINE 医学0002-9254AM J ENOL VITICULT AMERICAN JOURNAL OF ENOLOGY AND VITICULTURE农林科学0002-9262AM J EPIDEMIOL AMERICAN JOURNAL OF EPIDEMIOLOGY医学0195-7910AM J FOREN MED PATAMERICAN JOURNAL OF FORENSIC MEDICINEAND PATHOLOGY医学0002-9270AM J GASTROENTEROL AMERICAN JOURNAL OF GASTROENTEROLOGY医学1064-7481AM J GERIAT PSYCHIAMERICAN JOURNAL OF GERIATRICPSYCHIATRY医学1079-2082AM J HEALTH-SYST PAMERICAN JOURNAL OF HEALTH-SYSTEMPHARMACY医学0361-8609AM J HEMATOL AMERICAN JOURNAL OF HEMATOLOGY医学1049-9091AM J HOSP PALLIAT American Journal of Hospice andPalliative MedicineHEALTH CARE SCIENCES &SERVICES 1042-0533AM J HUM BIOLAMERICAN JOURNAL OF HUMAN BIOLOGY生物0002-9297AM J HUM GENET AMERICAN JOURNAL OF HUMAN GENETICS 生物0895-7061AM J HYPERTENS AMERICAN JOURNAL OF HYPERTENSION医学0271-3586AM J IND MEDAMERICAN JOURNAL OF INDUSTRIAL MEDICINE 医学0196-6553AM J INFECT CONTRO AMERICAN JOURNAL OF INFECTION CONTROL 医学0272-6386AM J KIDNEY DIS AMERICAN JOURNAL OF KIDNEY DISEASES 医学1088-0224AM J MANAG CARE AMERICAN JOURNAL OF MANAGED CARE 医学0002-9327AM J MATH AMERICAN JOURNAL OF 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american journal of gastroenterology杂志引文格式
(实用版)
目录
1.美国胃肠病学杂志(American Journal of Gastroenterology)简介
2.杂志的引文格式要求
3.引用示例
正文
美国胃肠病学杂志(American Journal of Gastroenterology)是一本知名的学术期刊,专注于发表关于胃肠病学和肝脏病学的研究文章。
该杂志创刊于 1919 年,由美国胃肠病学协会(American College of Gastroenterology)主办,是该领域研究的重要参考资料。
在引用美国胃肠病学杂志的文章时,需要遵循一定的引文格式。
本文主要介绍该杂志的引文格式要求。
一般来说,美国胃肠病学杂志的引文格式遵循以下原则:
- 作者姓名:文章作者的姓名应按照字母顺序排列,最多列出 7 位作者,超过 7 位作者时,需要使用“et al.”表示。
- 标题:文章标题应简短明了,准确反映文章主题。
- 期刊名称:需要使用全名,不能使用缩写。
- 出版年份:文章发表的年份,用小括号括起,紧跟在期刊名称后。
- 卷号和页码:文章所在的卷号和页码范围,用逗号分隔。
举个例子,如果您要引用一篇美国胃肠病学杂志的文章:
作者:张三,李四,王五等(共 7 位作者)
标题:关于胃肠病学的研究
期刊名称:美国胃肠病学杂志
出版年份:2021
卷号和页码:第 100 卷 (1-10 页)
那么引用该文章的格式应为:
张三,李四,王五等。
关于胃肠病学的研究。
美国胃肠病学杂志。
AMERICAN JOURNAL OF INDUSTRIAL MEDICINE38:355±360(2000) Relationship Between Delta-Aminolevulinic Acid Dehydratase Genotypes andHeme Precursors in Lead WorkersTadashi Sakai,PhD,1,2ÃYoko Morita,MD,DMSc,2Takaharu Araki,MD,DMSc,1,2 Mitsumasa Kano,MD,DMSc,3and Tsutomu Yoshida MD,DMSc4Background The objective of this study was to investigate the relationships betweengenotypes of delta-aminolevulinic acid(ALA)dehydratase(ALAD)and disturbances inthe heme biosynthetic pathway by lead exposure.Methods The subjects were192male lead workers and125control subjects.Blood leadconcentrations(Pb-B),plasma ALA concentrations(ALA-P),and ALAD genotypes weredetermined for all subjects.In lead workers,ALAD activity,ALA in urine(ALA-U),anderythrocyte zinc protoporphyrin(ZP)were also determined.Results The frequency of ALAD2(minor type of ALAD allele)was calculated to be0.087in all subjects.No signi®cant relationship was found between ALAD2frequency andPb-B levels in lead workers.ALAD1homozygotes showed signi®cantly higher levels ofZP and ALA-P in comparison with those of ALAD2carriers at Pb-B levels more than20m g/dL and40m g/dL,respectively.Conclusion ALAD1homozygotes might be more susceptible than ALAD2carriers todisturbances in heme metabolism caused by lead exposure.Am.J.Ind.Med.38:355±360,2000.ß2000Wiley-Liss,Inc.KEY WORDS:delta-aminolevulinic acid dehydratase;polymorphism;lead;delta-aminolevulinic acid;zinc protoporphyrin;plasma;urine;blood;susceptibilityINTRODUCTIONDelta-aminolevulinic acid(ALA)dehydratase(ALAD) is the second enzyme in the heme biosynthetic pathway which catalyses the condensation of two molecules of ALA to form one molecule of porphobilinogen[Granick and Mauserall,1958].Erythrocyte ALAD activity is rapidly inhibited by lead exposure[Lichtman and Feldman,1963] and as a result,ALA in plasma and urine(ALA-P and ALA-U)increases[Haeger-Aronsen,1971;Morita et al.,1994]. ALA is structurally similar to the neurotransmitter g-aminobutyric acid(GABA)and seems to compete with it for the receptor[Brennan and Cautrill,1979].Accumulated ALA may be responsible for the neurotoxic effects of lead in humans[Silbergeld and Lamon,1980].The®nal step of heme synthesis,introducing Fe2 into protoporphyrin IX (PP),is also affected by lead.Inhibition of Fe3 reduc-tion by lead exposure causes a decrease in transport of Fe2 into mitochondria[Taketani et al.,1985],resulting in an accumulation of PP in erythrocytes.PP is enzymatically or nonenzymatically chelated with Zn2 to form zinc proto-porpyrin(ZP).ALAD activity,ALA-P,ALA-U,and ZP are widely used as effect indices of lead exposure[Sakai and Morita,1996].In blood,more than98%of the lead is found in blood cells[DeSilva,1981;SchuÈtz et al.,1996].The amounts of plasma lead are relatively small,but it is very important1Center of Occupational Medicine,Tokyo Rosai Hospital,Tokyo,Japan2Industrial Poisoning Center,Tokyo Rosai Hospital,Tokyo,Japan3Center of Health Examination,Asahi Glass Corporation,Funabashi,Japan4Department of Public Health,Fujita Health University School of Medicine,Toyoake,Japan *Correspondence to:Tadashi Sakai,Centerof Occupational Medicine,Tokyo Rosai Hospital, 13-21,Omoriminami-4,Ota-ku,Tokyo,143-0013,Japan.E-mail:opc@msa.biglobe.ne.jp Accepted8March2000ß2000Wiley-Liss,Inc.because of its biological activity in lead metabolism.Lead in plasma circulates in the body,affects the body lead burden and causes the toxicity of lead in some soft tissues,such as bone marrow.The larger fraction of lead in blood cells is mainly bound to ALAD protein[Sakai et al.,1982;Bergdahl et al.,1997].Thus,ALAD might also play an important role in the dynamic interchange of the body lead pool.Human ALAD gene is located in chromosome9q34 [Potluri et al.,1987].The gene frequencies of ALAD alleles 1and2were reportedly about0.9and0.1,respectively, in several Caucasian populations[Battistuzzi et al.,1981; Benkmann et al.,1983;Astrin et al.,1987].In1991,a simple method using polymerase chain reaction(PCR)was developed for the analysis of the ALAD genotype[Wetmur et al.,1991a].In comparison with the ALAD1sequence, the only difference in the ALAD2cDNA was a G-to-C transversion of nucleotide177in the coding region,which creates an MspI restriction site and causes the replacement of a lysine by a asparagine[Wetmur et al.,1991a].The difference in the amino acid might result in a distinct charge isozyme,and the ALAD2protein may bind lead more tightly than a dose of the ALAD1protein[Wetmur,1994].Recently,several investigations have been conducted on the relationships between ALAD polymorphism and susceptibility to lead toxicity[Wetmur et al.,1991b; Schwartz et al.,1995,1997;Sakai et al.,1996b;Sithisas-rankul et al.1997;Alexander et al.,1998].In these studies, the effect indices of lead exposure,such as ALAD,ALA-P, ALA-U and ZP,were examined to determine whether the ALAD genotype in¯uences the values in the indices. However,none of the previous reports determined all the four indices in a suf®cient number of lead workers with a wide range of Pb-B levels.Furthermore,some disagree-ments were shown in the levels of Pb-B where differences in the effect indices existed between two different genetic groups.In the present study,we simultaneously examined four indices in suf®ciently large numbers of lead workers, and discussed the relationships between ALAD genotypes and the levels of the effect indices.MATERIALS AND METHODSVenous blood treated with heparin or EDTA-2K was obtained from192male lead workers.The lead workers were employed in a secondary smelter,two glass factories, in electrical appliance manufacture(soldering),or worked as painters.Venous blood treated with EDTA-2K was also obtained from125subjects(62males and63females)who had no occupational exposure to lead.These samples were collected during physical examinations.At the time of collection,spot urine was also collected from the lead workers.The study procedure was explained to all subjects and their informed consent was obtained.ALAD genotype was determined by the PCR-based method of Wetmur et al.[1991a].Pb-B was determined by ¯ameless atomic absorption spectrometry(Hitachi Z-8000, Tokyo,Japan).Erythrocyte ALAD activities were deter-mined by the Commission of European Communities(CEC) standard method,and the activity was expressed as units (u:m mol minÀ1L RBCÀ1)[Berlin and Schaller,1974].ALA in plasma and urine(ALA-P and ALA-U)was determined by the method of high-performance liquid chromatography (HPLC)previously reported[Morita et al.,1994;Sakai and Morita,1996a].ALA-U values were corrected for creatinine concentration.Creatinine concentration was determined by the method of Jaffe with the``Creatinine Determination Kit''by Wako Pure Chemicals(Osaka,Japan).ZP was determined by the HPLC method previously reported[Sakai et al.,1988].RESULTSTable I shows the distribution of ALAD genotypes and frequency of the ALAD2allele in the present study.ALAD2 frequency was calculated to be0.086in lead workers.In control subjects,ALAD2frequency was0.065in males and 0.111in females,though the difference between genders was not signi®cant(12test,12 1.33,P 0.247).In all control subjects,the ALAD2frequency was0.088,andTABLE I.Distribution ofALAD Genotypes and Frequency of the ALAD2Allele Among192Male Lead Workers and125Unexposed WorkersALAD genotypeWorkers1^11^22^2Total ALAD2frequencyLead(Male)1612921920.088Unexposed(All)1042011250.088(Male)5480620.065(Female)50121630.111Total2654933170.087356Sakai et al.almost equal to that in lead workers.Thus,no difference in ALAD2frequency was found between lead workers and control subjects.Table II shows the comparison of age,Pb-B,and ALA-P among ALAD genotypes in the control group.No signi®cant difference of age,Pb-B,or ALA-P was found between ALAD1homozygotes and ALAD2carriers for either gender.Table III shows the distribution of age,Pb-B,and indices of the heme biosynthetic pathway between two ALAD genotypes in lead workers.Means and ranges of age and Pb-B levels are nearly the same between the two genetic groups.In the heme parameters,levels of ALA-P and ZP in the ALAD1homozygotes are slightly higher than those in ALAD2carriers,although the difference is not signi®cant.Table IV shows the comparison of ALAD,ALA-P,ALA-U,and ZP levels between ALAD1homozygotes and ALAD2carriers in lead workers with different Pb-B levels.The workers were divided into ®ve groups according to Pb-B level.The mean value of each parameter was statistically compared by Student's t test or by Welch's test where their variance was signi®cantly different.The frequency of ALAD genotypes was not signi®cantly different among the ®ve groups.At Pb-B levels higher than 20m g/dL,ZP levels in ALAD1homozygotes are signi®cantly higher than that in ALAD2carriers.At Pb-B levels higher than 40m g/dL,ALAD1homozygotes have higher levels of ALA-P than those of ALAD2carriers.Figure 1(A)demonstrates the comparison of ALA-P levels between ALAD1homozygotes and ALAD2carriers in lead workers with increasing levels of Pb-B.The mean and standard deviation of ALA-P are plotted for the ®ve groups with various Pb-B levels.ALAD1homozygotes show signi®cantly higher levels of ALA compared with those of ALAD2carriers at Pb-B levels between 40andTABLE parison of Effect Indices of Lead Exposure Between Two ALAD Genotypes in Lead Workers (n 192)with Different Pb-B LevelsPb-B group (l g/dL)`2020^3940^5960^7980`Number of genotype 9139253318#6511Pb-B (m g/dL)7.7(5.0)28.2(5.9)49.1(6.6)67.4(7.1)86.3(8.0)8.3(4.8)27.2(6.3)49.1(6.4)61.290.0ALAD(u)57.8(14.4)30.0(9.6)14.6(5.2) 6.3(1.9) 5.5(1.9)53.8(12.1)28.7(5.8)14.9(3.0)9.5 5.5ALAP (m g/L)9.8(1.9)15.0(3.1)25.7(10.1)117.9(32.0)258.5(121.7)9.9(2.2)13.6(1.3)18.3(3.8)*53.1144.5ALAU (mg/gCre)0.9(0.4)1.1(0.4)1.9(1.0)8.7(4.2)26.4(9.9)0.9(0.3)1.1(0.4)1.5(0.5) 2.320.6ZP88(30)133(104)199(76)635(295)1319(555)(m g/dLRBC)73(23)79(34)*125(21)**291740Mean(SD):1stand 2ndrow:ALAD1homozygotes and ALAD2carriers.#:Containingtwosubjectswith ALAD2homozygotes.*P `0.05,**P `0.01compared with ALAD1homozygotes.TABLE II.Age and Concentration of Pb-B and ALA-P inTwo Genetic Groups of Control SubjectsMaleFemale Number ofgenotypes 5450813Age (years)41.3Æ9.1(20^59)22.3Æ5.1(19^40)42.3Æ7.8(34^56)24.7Æ9.9(18^50)Pb-B (m g/dL) 3.7Æ1.1(2.0^6.9) 2.4Æ0.8(1.4^3.7)3.6Æ1.0(2.5^4.9) 2.6Æ1.6(1.1^6.6)ALA-P (m g/L)9.0Æ2.1(6.0^15.7)7.5Æ1.5(5.0^10.9)9.2Æ2.2(6.4^11.0)6.6Æ1.4(5.1^8.7)Mean ÆSD (Range);1stand2ndrow:ALAD1homozygotes andALAD2carriers.TABLE III.Distribution of Age,Pb-B,and Indices of the Heme Biosynthetic Pathway BetweenTwo ALAD Genotypes in Lead WorkersALAD1homozygotesALAD2carriers (n 161)(n 31)Age (years)43.3Æ10.1(18^62)a 46.0Æ10.3(21^63)Pb-B (m g/dL)21.9Æ19.7(2.1^95.3)22.9Æ21.7(2.7^90)ALAD (u)42.5Æ22.1(4.1^81.8)39.7Æ20.1(5.5^74.9)ALA-P (m g/L)24.6Æ47.6(7.5^385.1)19.4Æ27.6(7.3^144.5)ALA-U (mg/gCre)1.5Æ3.5(0.2^31.1)1.6Æ3.6(0.4^20.6)ZP (m g/dLRBC)156Æ213(35^1684)117Æ134(38^739)aMean ÆSD (Range).ALAD Polymorphism and Heme Precursors 35759m g/dL.In lead workers with Pb-B levels higher than 60m g/dL,differences of ALA-P levels among the groups cannot be determined because the number of ALAD2carriers was as far as 2in certain groups.Figure 1(B)shows the comparison of ALA-U levels between the two genetic groups in lead workers.No signi®cant difference is observed between the two geno-types in every Pb-B groups,although ALAD1homozygotes show slightly higher levels than those of ALAD2carriers.Figure 1(C)shows the comparison of ZP levels between the two genetic groups in lead workers.ALAD1homo-zygotes show statistically higher levels of ZP in comparison with those in ALAD2carriers at Pb-B levels higher than 20m g/dL.These data also suggest that ZP is more sensitive to lead exposure than ALA in ALAD1homozygotes.DISCUSSIONIn the present study,the ALAD2frequency was calculated to be 0.087in all subjects (Table I).This frequency in the Japanese population is considerably different (12 3.56,P 0.059)from that by Benkmann et al.[1983](n 144,ALAD2frequency is 0.058).This discrepancy in ®ndings might be due to differences in the methods of determining the ALAD genotype or in the respective sample populations.Benkmann et al.[1983]used starch gel electrophoresis,which is based on the difference in the electrical charge of ALAD ing the PCR method,the frequency of the ALAD2allele has been reported in various populations were 0.114in 307Korean [Schwartz et al.,1995],0.074in 691American [Smith et al.,1995],and 0.085in 205Chinese [Zhang et al.,1998].These frequencies were nearly the same as those found in the present study.It has been reported that ALAD2carriers have higher Pb-B levels than ALAD1homozygotes in subjects exposed to lead occupationally or environmentally [Ziemsen et al.,1986;Astrin et al.,1987;Wetmur et al.,1991b;Schwartz et al.,1995].However,no signi®cant difference was observed in ALAD activity between the two groups ofALAD genotypes when their Pb-B levels were matched [Ziemsen et al.,1986].In the present study,ALAD activity is not signi®cantly different between the two ALAD genotypes in each Pb-B group (Table IV).In the present study,we demonstrated that ALAD genotypes modify lead effects on heme metabolism in highly exposed workers (Table IV ,Fig.1).On the other hand,in control subjects and lead workers with Pb-B levels lower than 20m g/dL,ALA and ZP levels were not signi®cantly different between the two ALAD genotypes (Tables II and IV).These data agree with those previously reported;ALAD genotypes were not found to affect the levels of heme precursors at low-Pb-B levels [Alexander et al.,1998;Zhang et al.,1998].At Pb-B levels of 20±59m g/dL,ZP levels in ALAD1homozygotes were signi®cantly higher than those in ALAD2carriers (Table IV ,Fig.1).At Pb-B levels of 40±59m g/dL,ALA-P levels in ALAD1homozygotes were also signi®cantly higher than those in ALAD2carriers (Table IV ,Fig.1).Sithisarankul et al.[1997]reported that ALA-P levels in ALAD1homozygotes were higher than those in ALAD2carriers in lead workers,whose results are in good agreement with ours.However,in the aforemen-tioned study,ZP levels were not signi®cantly different between the ALAD genotypes.One explanation for this difference may be the insuf®cient number (n 65)of workers in that study.In contrast,Schwartz et al.[1995]and Alexander et al.[1998]reported that ALAD1homozygotes had higher ZP levels than those of ALAD2carriers.In their studies,the number of subjects was suf®ciently large (n 308and 134,respectively),and ALA-P levels were not examined.The differences in the levels of heme precursors between the two types of ALAD genotypes might be attributable to a difference in the af®nity to lead among different ALAD isozymes ALAD1and two proteins.In a recent study using liquid chromatography in conjunction with inductively coupled plasma mass spectrometry,Bergdahl et al.[1997]reported that ALAD2carriers showed a higher percentage of lead bound to ALAD proteinthanparison(meanandSD)ofALA-P(A),ALA-U(B),andZP(C)levelswithincreasinglevelsofPb-Bbetweenthetwogenotypes.*P `0.05,**P `0.01.358Sakai et al.did ALAD1homozygotes.The amounts of``biologically active''lead might be lowered in ALAD2carriers than in ALAD1homozygotes.Thus,lead bound to the ALAD protein might be detoxi®ed or in inactive form,which prevents lead inhibition of ferrochelatase and of ferrous incorporation into mitochondria,resulting in relatively low levels of ZP in ALAD2carriers[Schwartz et al.,1995].The reductive process of Fe3+is reportedly inhibited by lead [Taketani et al.,1985].By these mechanisms,incorporation of ferrous into protoporphyrin might be more highly inhibited by lead in ALAD1homozygotes than in ALAD2 carriers,resulting in the®ndings of higher ZP and lower heme levels.Reduced levels of heme might induce ALA synthetase by negative feedback regulation,and produce a large amount of ALA[Meredith et al.,1978;Sakai et al., 1996a].This would explain the relatively higher levels of ALA in plasma in ALAD1homozygotes,in comparison to those in ALAD2carriers.ALA-U levels are not signi®cantly different between ALAD genotypes(see Fig.1(B)),which might be due to the fact that the urinary concentration and matrices are widely varied in each worker.In conclusion,ALAD1homozygotes have higher levels of ZP and ALA than those in ALAD2carriers in subjects who had high lead exposure,suggesting that ALAD1 homozygotes might be more susceptible than ALAD2 carriers to disturbance in heme biosynthesis caused by lead exposure.REFERENCESAlexander BH,Checkoway H,Costa-Mallen P,Faustman EM,Woods JS,Kelsey KT,van Netten C,Costa LG.1998.Interaction of blood lead and d-aminolevulinic acid dehydratase genotype on markers of heme synthesis and sperm production in lead smelter.Environ Health Perspect106:213±216.Astrin KH,Bishop DF,Wetmur JG,Kaul B,Davidow 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J.Dairy Sci.87:3561–3573American Dairy Science Association,2004.Guidelines for Monitoring Bulk Tank Milk Somatic Cell and Bacterial CountsB.M.Jayarao,S.R.Pillai,A.A.Sawant,D.R.Wolfgang,and N.V.HegdeDepartment of Veterinary Science,The Pennsylvania State University,University Park 16802ABSTRACTThis study was conducted to establish guidelines for monitoring bulk tank milk somatic cell count and bacte-rial counts,and to understand the relationship between different bacterial groups that occur in bulk tank milk.One hundred twenty-six dairy farms in 14counties of Pennsylvania participated,each providing one bulk tank milk sample every 15d for 2mo.The 4bulk tank milk samples from each farm were examined for bulk tank somatic cell count and bacterial counts including standard plate count,preliminary incubation count,laboratory pasteurization count,coagulase-negative staphylococcal count,environmental streptococcal count,coliform count,and gram-negative noncoliform count.The milk samples were also examined for pres-ence of Staphylococcus aureus ,Streptococcus agalac-tiae ,and Mycoplasma .The bacterial counts of 4bulk tank milk samples examined over an 8-wk period were averaged and expressed as mean bacterial count per milliliter.The study revealed that an increase in the frequency of isolation of Staphylococcus aureus and Streptococcus agalactiae was significantly associated with an increased bulk tank somatic cell count.Paired correlation analysis showed that there was low correla-tion between different bacterial counts.Bulk tank milk with low (<5000cfu/mL)standard plate count also had a significantly low level of mean bulk tank somatic cell count (<200,000cells/mL),preliminary incubation count (<10,000cfu/mL),laboratory pasteurization count (<100cfu/mL),coagulase-negative staphylococci and environmental streptococcal counts (<500cfu/mL),and noncoliform count (<200cfu/mL).Coliform count was less likely to be associated with somatic cell or other bacterial counts.Herd size and farm management practices had considerable influence on somatic cell and bacterial counts in bulk tank milk.Dairy herds that used automatic milking detachers,sand as bedding ma-Received June 5,2003.Accepted June 21,2004.Corresponding author:B.M.Jayarao;e-mail:bmj3@.3561terial,dip cups for teat dipping instead of spraying,and practiced pre-and postdipping had significantly lower bulk tank somatic cell and/or bacterial counts.In con-clusion,categorized bulk tank somatic cell and bacterial counts could serve as indicators and facilitate monitor-ing of herd udder health and milk quality.(Key words:bulk tank milk,somatic cell,bacterial count,milk quality)Abbreviation key:BTM =bulk tank milk,BTSCC =bulk tank somatic cell count,CC =coliform count,ES =environmental streptococci,LPC =laboratory pasteur-ization count,NC =noncoliform count,PIC =prelimi-nary incubation count,SA =Staphylococcus aureus ,SAG =Streptococcus agalactiae ,SPC =standard plate count.INTRODUCTIONSince the early 1990s,researchers have used bulk tank milk (BTM )to diagnose multiple problems (cur-rent and potential)that might exist in a dairy herd related to milk quality and mastitis pathogens.Progres-sive dairy producers,veterinarians,and dairy health consultants are interested in BTM analysis as a tool to determine milk quality and troubleshoot herds with mastitis.Many quality-conscious milk cooperatives have implemented BTM analysis to reward dairy pro-ducers who excel at producing high quality milk and have a low incidence of mastitis.In addition,milk pro-ducers and cooperatives view BTM analysis as an im-portant part of their quality assurance program (Emer-son,1989;Farnsworth,1993;Bray and Shearer,1996;Britten and Emerson,1996;Keeter,1997;Mickelson et al.,1998;Jayarao et al.,2001).Successful milk quality assurance programs start with farm BTM free of antibiotic residues and with low somatic cell and bacterial counts,resulting in better quality products with longer shelflife (Boor et al.,1998;Ma et al.,2000;Reugg and Tabone,2000).Many dairy producers also receive premiums from their milk coop-erative for producing milk with low somatic cell and bacterial counts.Several guidelines have been proposedJAYARAO ET AL. 3562to interpret BTM milk bacterial counts(Bray and Shearer,1996;Britt et al.,1997;Murphy,1997;Jones and Sumner,1999;Edmondson,2000;Jayarao et al., 2001;Jayarao and Wolfgang,2003).However,many of the guidelines are based on individual or collective experience,or extrapolations from other scientific stud-ies.Further,many of the interpretive guidelines lack validation and provide little insight into the interrela-tionship between different groups of bacteria found in BTM.An extension and research study,conducted in Pennsylvania from April2000through March2001, focused on BTM analysis.Thefindings of the milk qual-ity survey were used to establish guidelines for inter-preting BTM counts and also to understand the rela-tionship between different bacterial groups that occur in BTM.MATERIALS AND METHODSDairy HerdsThe veterinary extension group at Pennsylvania State University with the support of the county exten-sion agents implemented the study.A total of12county extension agents and1milk cooperative participated in the study.Each participating county extension agent/ milk cooperative enrolled7to11dairy producers from its county or region.Dairy producers who participated in the study were solicited by county extension agents through their extension newsletter or announcements about the study during a monthly dairy extension meet-ing.For a given county,participation in the study was open to all dairy producers,and thefirst12dairy pro-ducers who responded to the invitation were included in the study.Dairy producers who opted to participate in the pro-gram answered a self-administered questionnaire.The questionnaire sought information on the following as-pects of the dairy herd:1)herd size,2)milk production, 3)milking frequency,4)milkings per tank pickup,5) type of milking facility,6)change in milking facility, 7)use of automatic milking detachers,8)type(s)of bed-ding,9)animals purchased,10)residue violations in the past6mo,11)milk quality premiums in the past 6mo,12)type of milk equipment cleaning system,13) mastitis prevention and control practices,and14)milk-ing procedures.The questionnaire used in this study has been successfully used previously(Jayarao and Cassel,1999).The responses to the questions were ana-lyzed to determine if any of these practices were associ-ated with bulk tank somatic cell count(BTSCC)or bacterial counts.Journal of Dairy Science Vol.87,No.10,2004Collection and Processing of BTMThe county extension agent provided on-farm instruc-tion on BTM collection and handling procedures as de-scribed by National Mastitis Council BTM sample col-lection and handling guidelines(NMC,1999).Dairy producers collected the sample in thefirst and third week of each month for2mo(4samples total).Sampling kits containing gloves,racks,tubes(50mL sterile screw cap tubes),and labels were provided.Bulk tank milk samples were collected in sterile50-mL screw-cap cen-trifuge tubes.Within24hr of collection,all milk sam-ples were shipped on ice overnight to the laboratory. On receipt of the sample in the laboratory,only those samples that recorded a temperature of<7°C were pro-cessed.The BTM in the50-mL centrifuge tube was mixed thoroughly several times,and20mL of the milk was transferred to a snap-cap vial containing a preser-vative and sent to the Dairy One Laboratory in State College,PA,for determination of BTSCC.The remain-der of the milk sample was used for bacteriological analysis.Bacteriological Analysis of BTMThe BTM samples were examined for standard plate count(SPC),preliminary incubation count(PIC),labo-ratory pasteurization count(LPC),CNS count,environ-mental streptococci(ES)count,coliform count,and gram-negative noncoliform(NC)count.Bacteriological tests for milk quality were done as described by the American Public Health Association(Marshall,1992). The milk samples were mixed thoroughly by gently inverting the milk vial20to25times.One milliliter of milk was transferred to a sterile tube containing9mL of quarter-strength Ringer’s solution(Oxoid,Unipath Ltd.,UK).The10-fold diluted sample was vortexed at high speed for15s,and50µL was plated on selective and nonselective media using a spiroplater(Autoplate 4000,Spiral Biotech,Bethesda,MD).Plate count agar was used for enumeration of SPC,PIC,and LPC.The numbers of ES and Streptococcus agalactiae(SAG)in BTM samples were estimated using modified Edward’s agar supplemented with colistin sulfate and oxolinic acid(Sawant et al.,2002).MacConkey’s agar no.3(Ox-oid)was used to determine coliform and noncoliform counts.Baird Parker’s agar(Difco)was used to deter-mine the number of CNS and presence of Staphylococ-cus aureus(SA).Plates for enumeration of SPC,PIC, and LPC were incubated at32°C for48h.Plates for enumeration of CNS,ES,coliform count(CC),and NC were incubated at37°C for48h.The Autoplate4000 user guide(Spiral Biotech)was used to enumerate bac-terial counts.OUR INDUSTRY TODAY3563Colonies suggestive of SAG from modified Edward’s agar supplemented with colistin sulfate and oxolinic acid were randomly selected and streaked on 5%sheep blood agar and incubated for 48h at 37°C.All isolates were examined for gram’s reaction and catalase produc-tion,serotyped (Streptex,Oxoid),and identified usingAPI 20STREP (BioMe´rieux,Hazelwood,MO)(Sawant et al.,2002).Colonies suggestive of SA from Baird Par-ker agar were randomly selected,streaked on 5%sheep blood agar,and incubated for 48h at 37°C.The isolates were examined for hemolysis,catalase production,and coagulase production,and identified using API-STAPH (BioMerieux)(NMC,1999).Isolation of Mycoplasma was done as described by Gonzalez et al.(1995),with modifications.Briefly,500µL of BTM was pre-enriched in modified Hayflick’s broth and incubated for 48h at 37°C in a moist 10%CO 2incubator.One hundred microliters of the pre-enriched broth was streaked on modified Hayflick’s agar and incubated for 7d at 37°C in a moist 10%CO 2incubator.Mycoplasma colonies were viewed under a low-power microscope.Mycoplasma was differentiated from Acholesplasma laidlawdii using the digitonin inhibi-tion test as described by Thurmond et al.(1989).Data AnalysisA total of 149dairy herds elected to participate in the study of which 4herds opted not to participate during the course of the study.Of the 145herds,7dairy herds were unable to provide information on farm management practices,and 12dairy producers,on 2consecutive occasions,supplied contaminated bulk tank milk,or the milk that was received for analysis had a temperature in excess of 7°C.A total of 126dairy herds with complete data sets were used for data analysis.Answers to the questionnaire were transferred to Mi-crosoft Excel and grouped by their categorical response (e.g.,yes ,no ).To estimate if a response had an influence on the mean BTSCC,SPC,PIC,LPC,SA,CNS,ES,CC,and NC counts for each group within a response were compared with the 3categories (low,medium,high)within each bacterial count using one-way AN-OVA.A P -value of <0.05was considered a significant association between the response and a category of the count.All statistical analyses were performed using JMP software version 4.0(SAS Inst.,Inc.,Cary,NC).BTSCC and bacterial counts from the 4BTM samples from each farm were transformed to log10values.The log10transformed BTSCC and bacterial counts (SPC,PIC,LPC,SA,CNS,ES,CC,and NC)from the 4bulk tank samples from each farm were averaged and sub-jected to correlation coefficient analysis (SAS Inst.Inc.).Journal of Dairy Science Vol.87,No.10,2004The BTSCC,SPC,PIC,LPC,SA,CNS,ES,CC,and NC counts were each classified as low,medium,or high.These 3categories are the suggested interpretive crite-ria for monitoring BTM (see Table 2).The average counts for each of the 3groups were compared using the Tukey-Kramer (equal variance)or Dunnett’s T3(unequal variance)procedures.These 2procedures were used due to unequal sample sizes observed in the 3categories of a given count.The Tukey-Kramer procedure performs all pair-wise comparisons,testing whether the 3means are significantly different.The Dunnett’s T3procedure performs all comparisons with a control category.In our study,the second category (medium)was used as the control because the sample mean falls in this category.P <0.05was considered significant.Epi-info-2002(Centers for Disease Control and Prevention,Atlanta,GA),a database and statistics system for epidemiology on microcomputers,was used for performing χ2-square tests and odds ratio analysis.RESULTSDairy HerdsThe responses to the 14questions on the question-naire were grouped based on herd size (Table 1).Nearly 71%of the farms had fewer than 100lactating cattle,typical of farm families engaged in milking cows in Pennsylvania.Farm management practices changed as the herd size increased.This observation can be sup-ported by change in the management practices such as 1)number of milkings per day,2)type of milking facil-ity,3)use of automatic milking detachers,4)type of cow bedding,5)number of animals purchased,6)milk equipment cleaning system,7)mastitis prevention and control,and 8)milking practices.For the majority of the dairy herds,cows were milked twice a day (88%)in stanchion barns (61%)and/or parlors (39%).About 45%of dairy herds had automatic milking detachers.As the herd size increased,so did the use of automatic milking detachers.Sawdust was used as bedding on 44%of the farms surveyed.Nearly 5%of the respon-dents to the questionnaire indicated antibiotic residues in the last 6mo.A majority of the dairy producers practiced dry cow treatment (88%),whereas 73%of the dairy producers who teat dipped their cows practiced both pre-and king practices varied con-siderably within a given herd size and between the 4herd size categories (Table 1).Significant differences were observed with respect to the type of bedding used (P ≤0.000),antibiotic residues in bulk tank milk (P ≤0.043),type of milking equipment cleaning system (P ≤0.022),dry cow treatment (P ≤0.043),teat-dipping practices (P ≤0.031),stripping practices before milking (P ≤0.028),and towel type (P ≤0.011)(Table 1).JAYARAO ET AL.3564Table1.Characteristics of the dairy herds that participated in the study.Herd size<5050to99100to199>200Totalχ2Query(n=35)(n=55)(n=30)(n=6)(n=126)(P≤0.005) No.of cows in milk(average)386713629487Milk produced per cow(lb)3334333032.5Times milked(%)Two9791831788 4.99(0.111) Three39178312No.of milkings in bulk tank(%)Two010241711Three041004 6.63(0.011)* Four9782625080>Four343135Milking facilityStanchion9475100610.28(0.632) Parlor6259010039Change in milking facility in last6mo(%)1420303321 6.89(0.078) Automatic milking detachers(%)2652478345 4.54(0.122) Cow bedding(%)Combination(>1type bedding,c–i)119308Corn fodder34001Hay04002Mats3700411.86(0.000)* Newspaper611708Sand361707Sawdust3733638744Shavings141131310Straw23197016Animals purchased(%)Dry cows99303315Milking cows1116405022 2.70(0.145) Spring heifers2011335021Antibiotic residues in last6mo3940511.3(0.043)* Milk premiums in last6months7447556058 1.04(0.382) Milk equipment cleaning system(%)Automatic74969310088Manual320027.64(0.022)* Semi-automatic2327010Mastitis prevention and control(%)Dry cow therapy(always)8688931008811.37(0.043)* Teat dipping746787837314.54(0.031)* Predipping only37017612.01(0.007)* Postdipping only183310022Pre-and postdipping7960908372Milking practicesWritten protocols3971779.73(0.052) Check for mastitis38365880420.62(0.486) Wear gloves1812275019 4.96(0.112) Strip before milking636377836715.63(0.028)* Type of towelCommon wash cloth34704Individual wash cloth1118306721 6.69(0.011)* Paper towel7162533361Medicated towel151610014MilkersEmployees3217177Family members493120031 1.92(0.195) Self and employees920436725Self4037201737*P≤0.05.Bulk Tank Somatic Cell CountsThe mean BTSCC(315,190cells/mL)varied signifi-cantly with respect to the herd size.Fifty percent of the Journal of Dairy Science Vol.87,No.10,2004BTM samples had a BTSCC<348,000cells/mL.Paired correlation analysis showed that there was low correla-tion between BTSCC and different bacterial counts(Ta-ble2).Bulk tank somatic cell counts were categorizedOUR INDUSTRY TODAY3565T a b l e 2.D e s c r i p t i v e s t a t i s t i c s a n d c o r r e l a t i o n c o e f fic i e n t s o f c o u n t s o f b u l k t a n k m i l k (B T M )s a m p l e s f r o m 126d a i r y p r o d u c e r s i n P e n n s y l v a n i a .1M e a n c o u n tH e r d s i z eB T SC C 2S P C 3P I C 3L P C 3C N S 3E S 3C C 3N C 3<50320,440326091401507606303017050–99375,169476013,95015082090060210100–199289,175550012,22011054078080310>200283,895310037401005191010200130R a n g e 495,250–737,500180–62,820500–139,7505–6,40060–15,18015–1,10405–4,1300–15,460M e a n c o u n t (a l l h e r d s )4315,1904,320874012565082070200χ2(P <0.05)2.87(0.038)1.41(0.242)2.44(0.067)0.39(0.759)1.18(0.319)0.78(0.506)5.97(0.003)1.12(0.341)C u m u l a t i v e f r e q u e n c y <10%187,2501,1402,200301902101030<50%348,0004,21012,50013370090060230<90%553,25019,37062,0001,2402,6503,1002301,240C o r r e l a t i o n c o e f fic i e n t s B T S C C 10.320.1980.1480.3220.3620.180.108S P C —10.6190.510.5710.6480.3850.415P I C —10.5020.4350.5330.2390.435L P C ———10.3770.4050.1660.264C N S ————10.5030.1210.344E S —————10.1930.221C C ——————10.279N C———————11B T SC C =b u l k t a n k S C C ,S P C =s t a n d a r d c e l l p l a t e ,P I C =p r e l i m i n a r y i n c u b a t i o n c o u n t ,L P C =l a b o r a t o r y p a s t e u r i z a t i o n c o u n t ,C N S =c o a g u l a s e -n e g a t i v e s t a p h y l o c o c c i ,E S =e n v i r o n m e n t a l s t r e p t o c o c c i ,C C =c o l i f o r m c o u n t ,N C =n o n c o l i f o r m c o u n t .2c e l l s /m L (l o g t r a n s f o r m e d v a l u e s ).3c f u /m L (l o g t r a n s f o r m e d v a l u e s ).4126d a i r y h e r d s .Journal of Dairy Science Vol.87,No.10,2004JAYARAO ET AL.3566Table 3.Categorization of mean bulk tank somatic cell and bacterial counts.1Proposed interpretive criteriaBulk tank Category Count (cfu/mL)N 2BTSCC SPC PIC LPC CNS ES CC NC Low <20019179,3902290654090360*39030120BTSCC Medium 200,000–400,00055283,320*41401044013068076060290High ≥400,00052497,310597014960160940108070220Low <5,00070310,90019506170*80*440*490*40130*SPC Medium 5,000–10,00024303,6017470*168702801170122070350High ≥10,00032415,04017680312902801410169090470Low <10,00060313,6102370*366080470*460*40120PIC Medium 10,000–20,00020337,640510013290*1301030117060200High ≥20,00026358,110928044230290*1040135070440Low <10052307,86027206130*3247053050140LPC Medium 100–20021313,700290011870140*63078040270High ≥20053368,4308340*201405401120*1190*80280Low <50046277,520*249058108026045050120CNS Medium 500–100040350,140369010370*110720*82050180High ≥100040390,76010140*26400290*21601470*80460Low <50033284,9601940509060*38019041140ES Medium 500–100035311,4903480*10110*150540690*60170High ≥100058378,9708090189702001190*197070310Low <5057307,9903130852011060062020140CC Medium 50–10028356,160498011630160103095070*290High ≥10041354,710651016330170700980220290NCLow <20053303,5702980*67901005606504060Medium 200–40044359,24047701264014066079060270*High≥40029351,750797024030*240122011409013001See Table 2for abbreviation definitions.2N,number of bulk tanks *P ≤0.05.into 3groups (low,<200,000;medium,200,000to 400,000;and high,>400,000cells/mL)(Table 3).Mean CNS count was significantly associated with mean BTSCC (Table 3).A BTM with a mean BTSCC >200,000cells/mL was 5times more likely to have high CNS (>500cfu/mL)counts compared with BTM with BTSCC <200,000cells/mL (Table 4).Dairy producers who re-ceived milk premiums had significantly lower BTSCC (291,300cfu/mL)compared with the BTSCC (378,090cells/mL)in BTM of those dairy producers who did not receive premiums [χ2(p )=3.27(0.0014)].BTSCC was significantly lower when cows were milked using auto-matic milk detachers as compared with BTM from herds that milked cows without automatic milk detach-ers.The same observation was made with herds thatTable 4.Odds ratio (confidence interval)estimates for somatic cell and bacterial counts.1BTSCCSPCPICLPCCNSCounts>200,000cells/mL >5,000cfu/mL >10,000cfu/mL >100cfu/mL >500cfu/mL BTSCC >200,000cells/mL —————SPC >5,000cfu/mL —————PIC >10,000cfu/mL —9.55(3.86–24.15)———LPC >100cfu/mL —4.89(2.07–11.75) 3.02(1.36–6.77)——CNS >500cfu/mL 5.04(2.11–12.97) 5.86(2.32–15.12) 3.13(1.37–7.17) 3.71(1.56–8.94)—ES >500cfu/mL — 6.80(2.23–22.12) 4.22(1.64–11.12) 2.93(1.20–7.23) 5.75(2.25–14.94)NC >200cfu/mL—6.14(2.54–15.12)3.73(1.66–8.47)——1See Table 2for abbreviation definitions.Journal of Dairy Science Vol.87,No.10,2004teat dipped the cows with a dip cup instead of using a spray.Interestingly,BTSCC was significantly higher in herds that practiced fore-stripping before milking compared with BTM from herds that did not.Dairy farms that used sand as bedding had significantly lower BTSCC in their BTM compared with dairy producers who used organic bedding such as shavings,newspaper,and straw (Table 5).Standard Plate CountFor the 126dairy herds in the study,the mean SPC for an 8-wk period was 4320cfu/mL.The herd size did not influence the mean SPC of BTM.Fifty percent of BTM samples had a SPC <4120cfu/mL.Paired correla-OUR INDUSTRY TODAY3567Table 5.Effect of herd size and management practices on somatic cell and bacterial counts.1Herd sizePracticeCount<5050–99100–199>200Total Automatic Yes 354,580289,430*230,890*473,250298,560*detachers No 363,780356,000421,720—352,650Strip Yes 409,030*327,510341,070390,020357,060No 293,050295,570253,470313,750295,760*SprayYes 515,070295,580399,480361,370396,600No 335,840*327,510284,050459,500321,550*SCCDip cupYes 317,480*296,950300,710361,700306,360*No429,510359,450336,810459,500370,090Combination 295,450286,420306,250—291,940Corn fodder 449,330———449,330Hay —242,510——242,510Mats587,000342,760——381,700BeddingNewspaper 479,300343,490382,090—325,480Sand 316,750206,290*179,550*—241,750*Sawdust 350,320299,020343,550—317,990Shavings 315,420339,270274,670370,150353,580Straw 397,140413,200588,180407,500360,700Pre and postYes 358,030330,680282,800*408,340312,290*SPCNo 368,820284,310441,290319,190380,680SprayYes 94505100633062006710No 469043302430*21203910*PIC Spray Yes 26,03023,52015,90012,99018,300No 14,21010,1006200*16,20010,170*CNS Automatic Yes 660610400*1880510*detachers No 1110610870—860ESPre and postYes 900420*540520670*No1390930830133011201See Table 2for abbreviation definitions.*P ≤0.05.tion analyses between SPC and other bacterial counts showed that SPC had correlation coefficients >0.5for ES (0.648),PIC (0.618),CNS (0.571),and LPC (0.510)(Table 1).SPC were categorized into 3groups (low,<5000;medium,5000to 10,000;and high,>10,000cells/mL)(Table 3).The mean PIC,LPC,CNS,ES,and NC counts were significantly different for the 3SPC catego-ries (low,medium,and high)(Table 3).BTM with a mean SPC >5000cfu/mL were 9.5,5,6,7,and 6times more likely to have medium or high PIC,LPC,CNS,LPC,ES,and NC,respectively,compared with BTM with SPC <5,000cfu/mL (Table 4).The SPC was sig-nificantly lower in BTM when cows were subjected to both pre-and postdipping.In contrast,BTM samples had significantly higher SPC when cows were sprayed with a teat dip instead of using a dip cup (Table 5).Preliminary Incubation CountThe mean PIC for an 8-wk period ranged from 500–139,750cfu/mL with a mean PIC of 8740cfu/mL.As observed with SPC,herd size did not influence the mean PIC of BTM.Nearly 50%of the BTM milk samples had a PIC of <12,500cfu/mL.In addition to SPC,PIC had correlation coefficients >0.5for LPC (0.501)and ES (0.533)(Table 2).The mean SPC,LPC,CNS,and ESJournal of Dairy Science Vol.87,No.10,2004counts were significantly different for the 3PIC catego-ries (low,medium,and high)(Table 3).The BTM with a mean PIC >10,000cfu/mL were 3,3,4,and 4times more likely to have medium or high LPC,CNS,ES,and NC,respectively,compared with BTM with PIC <10,000cfu/mL (Table 4).As observed with SPC,BTM had significantly higher PIC when cows were sprayed with a teat dip instead of using a dip cup (Table 5).Laboratory Pasteurization CountThe mean LPC for an 8-wk period was 125cfu/mL.The herd size did not influence the LPC of BTM.Approx-imately 10%of BTM samples had a LPC <30cfu/mL,whereas 90%of the BTM samples had a LPC of <1240cfu/mL (Table 2).The mean SPC,PIC,CNS,and ES counts were significantly different for the 3LPC catego-ries (low,medium,and high)(Table 3).The BTM with a mean LPC >100cfu/mL were 4and 3times more likely to have medium or high CNS and ES,respectively,compared with BTM with LPC <100cfu/mL (Table 4).None of the management practices had any significant effect on LPC in BTM (Table 5).Coagulase-Negative StaphylococciThe mean CNS for an 8-wk period ranged from 60to 15,180cfu/mL with a mean CNS of 650cfu/mL.AsJAYARAO ET AL. 3568observed with other bacterial counts,herd size did not have any significant effect on CNS count of BTM.Fifty percent of BTM samples had CNS counts of<700cfu/ mL(Table2).The mean BTSCC,SPC,PIC,LPC,and ES counts were significantly different for the3CNS categories(Table3).The BTM samples with a mean CNS>500cfu/mL were6times more likely to have medium or high ES compared with BTM with CNS<500 cfu/mL(Table4).The CNS counts were significantly lower in BTM when cows were milked using automatic milk detachers compared with BTM from cows that were milked without the use of automatic milk detach-ers(Table5).Environmental StreptococciThe mean ES for an8-wk period was820cfu/mL. The herd size did not have a significant effect on ES count of BTM.Fifty percent of BTM samples had ES counts of<900cfu/mL.Ten and90%of the BTM samples had ES counts of<210and<3100cfu/mL,respectively. Paired correlation analyses between ES and CNS had a correlation coefficient of0.503(Table2).The mean SPC,PIC,LPC,and CNS counts were significantly dif-ferent for the3ES categories(Table3).The ES count was significantly lower in BTM when cows were pre-and postdipped(Table5).Coliform CountThe mean CC for an8-wk period was70cfu/mL.A significant association was observed between CC and herd size.As the herd size increased,so did the CC of BTM.About50%of BTM samples had CC<60cfu/mL (Table2).There was no significant difference in the mean of all of the bacterial counts for the3CC catego-ries(low,medium,and high)(Table3).Gram-Negative Noncoliform BacteriaFor the126dairy herds,the mean NC count was200 cfu/mL.As observed with other bacterial counts,herd size did not have any significant effect on an NC count of BTM.About50%of the BTM samples had an NC count of<230cfu/mL(Table2).The mean SPC and PIC were significantly different for the3NC categories(low, medium,and high)(Table3).The BTM samples with a mean NC>200cfu/mL were6and4times more likely to have medium or high SPC and PIC counts compared with BTM with NC<200cfu/mL(Table4). Contagious Mastitis PathogensThe bulk tank milk that tested positive for contagious mastitis pathogens was categorized as low frequency Journal of Dairy Science Vol.87,No.10,2004(1of4samples positive),medium frequency(2of4 samples positive),or high frequency(3or4samples positive)(Table6).Based on the analysis of4milk samples from each bulk tank,SA was detected in39of 126(31%)bulk tanks.It was observed that17,8,and 6%of the BTM samples had low,medium,and high isolation rates of SA,respectively.As the frequency of sampling increased from2to4samples,the number of bulk tanks with SA also increased.The mean BTSCC count was significantly associated with the frequency of isolation of SA(Table6).Streptococcus agalactiae was detected at least once in13of126(10%)bulk tanks (Table6).Of the BTM samples,3,5,and2%had low, medium,and high isolation rates of SAG,respectively. As seen with SA,with increased frequency of sampling, the number of BTM samples with SAG was also ob-served(Table6).Mycoplasma was isolated from3of39 (7.5%)BTM samples examined.DISCUSSIONFarnsworth(1993)presented thefirst set of guide-lines for interpreting BTM counts.This was followed by Bray and Shearer(1996),who developed comprehen-sive interpretive criteria for BTM counts.Murphy (1997)suggested interpretive guidelines for monitoring BTM counts,focusing on milk and milking system hy-giene.Other researchers have also provided guidelines for monitoring bulk tank bacterial counts as they relate to herd udder health and milk quality,with an empha-sis on troubleshooting herds with high bacterial counts (Britt et al.,1997;Jones and Sumner,1999;Edmond-son,2000;Jayarao et al.,2001;Jayarao and Wolfgang, 2003).These recommended guidelines served as the foundation for developing interpretive guidelines for monitoring BTM(Table3).Based on the1996to1997BTSCC data collected from dairy herds from49states,Pennsylvania ranked20th, with a state average of331,000cells/mL of milk(Nor-man et al.,2000).Paired correlation analyses between BTSCC and bacterial counts showed low correlations (Table1).Van Schaik et al.(2002),studying the trends of somatic cell counts in New York State during1999 to2000,observed that the average BTSCC was363,000 cells/mL.Thefindings of their study suggest that larger farms had lower BTSCC and plate loop count,but had more antibiotic residue violations.An individual cow somatic cell count of<200,000cells/ mL is typical of an uninfected udder(Laevens et al., 1997).A similar guideline was used by Van Schaik et al.(2002)for evaluating trends in somatic cell counts in New York.In our study,15%of the BTM had SCC <200,000cells/mL.This suggests that lowering BTSCC is still a challenge for many dairy producers in Pennsyl-。