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Biomedical subjects

J C Forrest

Publications and source records attributed to J C Forrest.

At least 19 recordsLinked to original sources

Pork carcass composition derived from a neural network model of electromagnetic scans.

We used an advanced computer logic system (NETS 3.0) to decipher electromagnetic (EM) scans in lieu of traditional linear regression for estimation of pork carcass composition. Fifty EM scans of pork carcasses were obtained on-line (prerigor) at a swine slaughter facility. Right sides were cut into wholesale parts and dissected into fat, lean, and bone to obtain total dissected carcass and primal cut lean. In this study, the input layer consisted of 81 nodes (80-point EM scan curve and warm carcass weight), one hidden layer of 42 nodes, and an output layer consisting of one node, which were run separately for outputs of ham, loin, or shoulder lean. The hidden layer connected to the output of total lean contained 50 nodes. Thirty-five scans were used for training of the network. The new network was then tested with 15 previously unseen input/output pairs. Separate neural networks were developed for the estimation of dissected total carcass, ham, loin, and shoulder lean. The NETS configuration improved on linear regression equations for estimation of total carcass lean by .31 kg, ham lean by .284 kg, and shoulder lean by .148 kg. Our results show that advanced computer logic systems have the capacity to improve upon traditional linear regression equations for prediction of pork carcass composition.

Animals

Evaluation of electronic technology to assess lamb carcass composition.

Accurate price signals are essential for producers of American lamb to ensure production of uniformly lean animals. Development of carcass merit-pricing systems will require the use of objective technology for assessing carcass composition or lean distribution. The objective of this study was to evaluate electronic technologies for accurate determination of lamb carcass composition. Lambs (n = 106) were selected as a representation of U.S. market lambs that transcended geographic location, sex, breed, carcass weight, yield grade, and production system. The independent variables used to predict lamb composition varied with the technology. The electronic technologies tested included realtime ultrasound, optical reflectance probe, bioelectrical impedance analysis, and electromagnetic scanning (TOBEC). All technologies, except realtime ultrasound, were tested on warm (prerigor) carcasses and repeated after a 24-h chill. Longitudinal ultrasonic scans of fat and muscle tissue depth and grading probe fat depths were marginal predictors of proportional carcass yield. The TOBEC measurements often accounted for more variability associated with kilograms of dissected lean and percentage of carcass lean than did carcass weight. Equations from TOBEC measurements were the most accurate predictors of weight and percentage of dissected and fat-free lean. Bioelectrical impedance measurements of resistance and reactance combined with carcass weight were also good predictors of carcass composition. Prediction of carcass lean distribution by measures of TOBEC were the most accurate for prediction of leg lean. The implications of usefulness of these technologies will depend on the commitment of the U. S. sheep industry in development of a lamb price discovery system based on carcass composition.

Animals

Assessment of lamb carcass composition from live animal measurement of bioelectrical impedance or ultrasonic tissue depths.

Market weight lambs, average weight 52.5 kg (+/-6.1), were used to evaluate nontraditional live animal measurements as predictors of carcass composition. The sample population (n = 106) represented U.S. market lambs and transcended geographic location, breed, carcass weight, yield grade, and production system. Realtime ultrasonic (RU) measurements and bioelectrical impedance analysis (BIA) were used for development and evaluation of prediction equations for % boneless, closely trimmed primal cuts (BCTPC), weight or % of dissected lean tissue (TDL), and chemically derived weight or % fat-free lean (FFL). Longitudinal ultrasonic images were obtained parallel to the longissimus thoracis et lumborum (LTL), positioning the last costae in the center of the transducer head. Images were saved and fat and LTL depths were derived from printed images of the ultrasonic scans. Bioelectrical impedance analysis was administered via a four-terminal impedance plethysmograph operating at 800 microA at 50 kHz. Impedance measurements of whole-body resistance and reactance were recorded. Prediction equations including common linear measurements of live weight, heart girth, hindsaddle length, and shoulder height were also evaluated. All measurements were taken just before slaughter. Bioelectrical impedance measurements (as compared to RU and linear measurements) provided equations for %BCTPC, TDL, %TDL, FFL and %FFL with the highest R2 and lowest root mean square error. Even though BIA provided the best equations of the three methodologies tested, prediction of proportional yield (%BCTPC, %TDL, and %FFL) was marginal (R2 = .296, .551, and .551, respectively). Equations combining BIA, RU, and linear measurements greatly improved equations for prediction of proportional lean yield.

Animals

Electrical measurement for detecting early postmortem changes in porcine muscle.

Use of electrical measurements to detect quality defects in porcine muscle in the early postmortem period was evaluated. Justification for use of a tetrapolar, constant current electrode configuration instead of bipolar electrodes was provided for measurements at low frequencies. Interrelationships among electrical properties, pH values, ATP decline, temperature, time postmortem, and final water-holding capacity (WHC) of porcine muscle were quantified using 25 hogs. Immediately after exsanguination, a section of the left longissimus muscle (LM) was excised to obtain rigor shortening patterns and complex impedance measurements over a 10-h period at 37 degrees C. Complex impedance measurements were taken using a tetrapolar electrode configuration at 1 kHz and .156 mA. At 15, 45, and 90 min postmortem, pH, ATP/IMP absorbance (R), and conductivity measured by the Tecpro Pork Quality Meter (PQM) were measured on the right side LM. At 24 h postmortem, WHC, pH, R, PQM, Hunter Color Lab values, and subjective quality scores were evaluated on the left LM. The WHC measurements were used to group carcasses into normal (n = 17) and abnormal (n = 8) categories. Mean pH and R at 45 and 90 min were different (P < .05) but pH at 24 h was not different between the normal and abnormal groups. Onset and completion of rigor were more rapid in carcasses with low WHC (P < .05). The PQM values were greater (P < .05) in the abnormal group at 90 min and 24 h postmortem. Excised muscle measurements of relative impedance (Z*) and phase (theta*) showed Z* and theta* increased more rapidly within the first 15 min postmortem (P < .1) for samples with abnormal WHC. However, one PSE carcass showed an immediate rapid decrease in Z* and theta*. Results suggest measurement of rate of change of impedance and phase angle before 90 min postmortem would be a better prediction of ultimate quality than absolute magnitude of impedance.

Adenosine Triphosphate

Alternative pork carcass evaluation techniques: I. Differences in predictions of value.

Dissected and predicted wholesale and lean boneless values for 154 pork carcasses representing seven genotypes with substantial variation in carcass composition and percentage of lean were determined. Dissected carcass value was determined using a component pricing model, and four alternative models were specified to predict that value. The models included measurements from a ruler (RULER) and two carcass evaluation technologies, Hennessy probe (PROBE) and electromagnetic scanner (EMS1). A combination of the PROBE and EMS1 models (EMS2) was also used. For wholesale value, R2 were .40, .70, .59, and .74, and the RSD were 8.18, 5.77, 6.76, and 5.38 ($/100 kg of carcass value) for RULER, PROBE, EMS1, and EMS2, respectively. For lean boneless value, the R2 were .41, .73, .59, and .74, and the RSD were 8.34, 5.67, 6.99, and 5.51 ($/100 kg of carcass value) for RULER, PROBE, EMS1, and EMS2, respectively. The results indicate that a combination of probe and electromagnetic scanner measurements provided the best fit to dissected value.

Animals

Alternative pork carcass evaluation techniques: II. Statistical analysis of error attributable to sex, genotype, and weight.

Carcasses of 154 hogs representing seven genotypes with substantial variation in carcass composition and percentage of lean were completely dissected and analyzed. Measurements from a ruler, Hennessy probe, and electromagnetic scanner were each used to predict wholesale and lean boneless carcass value. Error, defined as dissected value minus predicted value, due to the omission of sex, genotype, weight, and their interactions was estimated for each model. The errors were significantly different from zero for the models using ruler and electromagnetic scanning measurements separately (P < .01). Errors due to sex, genotype, weight, and their interactions were greatest for the less lean barrows. A combination of probe and electromagnetic scanner measurements resulted in the least error. The value of barrows with low percentage of lean was consistently overpredicted, whereas the value of leaner gilts was underpredicted for the models using ruler and electromagnetic scanning separately (P < .001).

Analysis of Variance

Electromagnetic scanning of pork carcasses in an on-line industrial configuration.

The objective of this study was to test and validate electromagnetic scanning of whole pork carcasses in an on-line, integrated, industrial configuration. The electromagnetic (EM) scanner was installed in two pork processing facilities (Plant A and Plant B). Plant A was a small pork fabrication plant that further processed chilled pork carcasses. Carcasses were delivered to Plant A by refrigerated trucks. The amount of EM energy absorbed by the carcasses was recorded as they were conveyed through the EM field. A plot of the absorption units over time (EM scan curve) was used to obtain predictive variables for estimating carcass and primal cut composition. Forty-eight whole, chilled carcasses (Group A) were electromagnetically scanned and conveyed onto the fabrication line. The average percentage carcass lean for Group A was 49.1% (range = 36.5 to 59.5%). Right carcass sides were removed from the processing line, fabricated into primal cuts, and dissected into fat, lean, and bone. Prediction equations were developed from EM scans for weight of total dissected carcass lean (R2 = .830; root mean square error = 1.80 kg), percentage of carcass lean (R2 = .820; root mean square error = 2.29%), and weight of dissected ham, longissimus muscle, and shoulder lean. In Plant B, the electromagnetic scanner was installed at the end of a pork slaughter line to ensure carcass scanning at a consistent carcass temperature. Fifty whole, pre-rigor eviscerated carcasses (Group B) were electromagnetically scanned before entering the chill cooler where fat and loin tissue depths were obtained by an optical grading probe. The average percentage carcass lean for Group B was 46.7% (range = 30.1 to 57.3%). Prediction equations were developed from EM scans for weight of total dissected carcass lean (R2 = .904; root mean square error = 1.59 kg), percentage of carcass lean (R2 = .863; root mean square error = 2.05%), and weight of dissected ham, loin, and shoulder lean. Statistical equations developed for the prediction of dissected primal cut lean were superior from EM scans of Group B (prerigor) carcasses. Electromagnetic scanning proved more statistically efficient than optical probes for predicting weight of dissected carcass lean and percentage of carcass lean. Statistical comparison of EM scan equations from Groups A and B are not completely valid because two different populations of carcasses were tested at different times of the year. The results of this study show that EM scanning has the potential to accurately predict pork carcass composition in a fully automated, on-line industrial configuration.

Animals

Electromagnetic scanning of beef quarters to predict carcass and primal lean content.

To study the use of electromagnetic scanning in prediction of lean content in beef carcasses and cuts, 100 beef cattle (60 steers and 40 heifers), representing a broad range in external fat thickness (.1 to 2.9 cm) and live weight (414 to 742 kg), were selected. Chilled right sides were divided into streamlined (foreshank, brisket, and ventral plate removed) forequarters (FQ) and full hindquarters (HQ) and scanned. Primal rounds, loins, ribs, and chucks were fabricated from the right side, scanned, and physically separated into lean, fat, and bone. Prediction equations for dissected lean content and percentage of lean included the peak of the electromagnetic scan response curve (obtained from scanning the HQ or FQ), length, temperature and weight of the scanned cut, and fat thickness at the 12th rib. Using the coefficient of determination, root mean square error, and Mallows' Cp statistic, the best model for each dependent variable (weight and percentage of lean) that included up to five independent variables was selected. Prediction equations for the HQ or FQ of steers accounted for 84 to 93% of the variation in lean weight of beef sides and quarters and 71 to 93% of primals. Sixty-one to 75% of the variation in percentage of lean in sides and quarters and 48 to 65% of primals was also explained. Similar results were obtained for heifer carcasses. Predicting percentage of lean in any scanned cut, rather than weight of lean, accounted for less of the variation. Weight and fat thickness contributed significantly when predicting percentage of lean.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Electromagnetic scanning to predict lamb carcass composition.

The electromagnetic scanner generates a constant, low-level electromagnetic field (2.5 MHz) within a large plexiglass tube. The amount of electromagnetic (EM) energy transferred (to the carcass) is highly related to lean tissue. A plot of the absorption units over distance can be used to assess the total mass of lean tissue and of the respective primal cuts. The difference in curve height between two points (D), peak phase absorption, and linear carcass measurements (pre-rigor, HCWT or post-rigor, CWT carcass weight, and carcass length, LENG) were used to predict total dissected lean (TOTLEAN), dissected leg lean (LEGLEAN), and percentage of dissected carcass lean (PERLEAN). Twenty-one pre-rigor and 22 post-rigor (24 h chill) lamb carcasses, average weight 26.8 (+/- 4.2 kg) and 26.4 (+/- 4.1 kg) kg, respectively, were evaluated from measurements of total body electrical conductivity (TOBEC). Two geometric orientations were tested for statistical accuracy in this study: A) each carcass entered the EM tunnel rear leg first, on its left lateral side, neck facing the right side of the tunnel; and B) each carcass entered the EM tunnel rear leg first, breast down, and neck up. Orientation A proved more statistically efficient for pre-rigor carcasses, and orientation B was more desirable for post-rigor carcasses. Multiple-regression models involving HCWT, LENG, and a single D measure accounted for 98.0 and 95.0%, respectively, of the total variation in pre-rigor carcass TOTLEAN and LEGLEAN in A.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Palatability of prerigor cooked boar meat.

Cooking reduces odor intensity in boar meat but also may induce lipid oxidation unless the meat pH is above approximately 6.0. This research was designed to determine the feasibility of cooking boar meat in the prerigor state to overcome boar odor and lipid oxidation problems. Prerigor and postrigor triceps brachii muscle samples from 40 boars (20 Duroc and 20 Yorkshire) were cooked to 60 degrees C, frozen and stored at -20 degrees C, reheated in a 60 degrees C water bath for 1 h, and then subjected to pH, thiobarbituric acid (TBA), and sensory analyses. Boar odor intensity and skatole concentration in backfat samples were determined by olfactory test and HPLC, respectively. Cooked (initial cooking) prerigor meat was found to have higher (P < .05) pH and lower (P < .05) TBA values than comparable postrigor meat (6.44 vs 6.09 and 2.15 vs 3.23, respectively). Regression analysis indicated an inverse relationship between pH and TBA values (r = -.52; P < .01). No appreciable changes in TBA values were noted after frozen storage for 14 to 98 d, but reheating increased TBA values (P < .05) in both prerigor and postrigor samples (3.45 vs 4.32, respectively). Sensory evaluation scores indicated that prerigor cooked meat was less tender with more pronounced rancid flavor than postrigor cooked meat (P < .05), but panelists may have allowed the toughness of the prerigor samples to adversely affect their flavor scores. No difference in boar odor was detected between rigor states or breeds. Mean skatole concentration in backfat was .12 micrograms/g and no difference was detected between breeds.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue

Effect of heating rate on palatability and associated properties of pre- and postrigor muscle.

Precooked, uncured meat is not widely available to consumers, partially because of associated palatability problems and lack of published information on heat uptake under different industrial conditions. The objectives of this study were to determine the tenderness, extent of lipid oxidation, and total cooking losses in pre- and posterior beef and pork roasts heated at different rates. The muscles were cooked in stainless-steel, perforated heating chambers at oven temperatures of 150, 200, or 250 degrees C and the temperature rise during and after heating was monitored with a digital temperature recorder. Samples were vacuum-packaged, frozen at -20 degrees C for 45 d, thawed at 4 degrees C for 24 h, and reheated in 60 degrees C water for 1 h. Cooking losses, Warner-Bratzler shear force values, thiobarbituric acid values, and pH were determined. The results provide heating curves for pre- and postrigor beef and pork roasts at three oven temperatures. Prerigor samples of both species were less tender than postrigor samples (P < .05). Cooking losses were generally low in prerigor samples of both species compared with postrigor samples (P < .05). All beef samples had relatively low thiobarbituric acid (TBA) values before and after storage, whereas pork samples had relatively high TBA values before and after storage. Results indicate that prerigor cooked roasts shrink less, are equivalent or better in oxidative stability, and are less tender than postrigor cooked roasts under the conditions of this experiment.

Animals

Evaluation of alternative techniques to determine pork carcass value.

Three techniques for estimating the value of pork carcasses were evaluated: an optical probe, a real-time ultrasound scanner, and an electromagnetic scanner (EMSCAN). The ability of these techniques to predict carcass value was compared to the predictive ability of actual measures of backfat depth and longissimus muscle area taken with a ruler and a dot grid. Results indicated the EMSCAN model was the best predictor of carcass value. However, the optical probe, ultrasound, and the ruler/dot grid all provided information not contained in the EMSCAN model. The choice among ultrasound, the optical probe, and the ruler/dot grid depends on how the carcass will be used. There is no significant difference between ultrasound and the ruler/dot grid or the optical probe and the ruler/dot grid if the carcass is to be marketed in wholesale primal form, but the ruler/dot grid is superior if the ham and loin are to be sold as lean, boneless products. A model combining the EMSCAN and optical probe readings provided more accurate value predictions than either technique alone. A carcass value matrix for use in pricing pork carcasses was developed using readings from the optical probe. Carcass use has a substantial impact on value differences between fat and lean pigs.

Adipose Tissue

Genotype and treatment biases in estimation of carcass lean of swine.

Carcasses of 181 barrows, representing five genotypes, 1) H x HD, 2) SYN, 3) HD x L[YD], 4) L x YD, and 5) Y x L (H = Hampshire, D = Duroc, SYN = synthetic terminal sire line, L = Landrace, and Y = Yorkshire), and two levels of ractopamine (RAC) treatment (0 and 20 ppm) were completely dissected and the data were used to examine genotype and treatment (RAC) biases in estimation of fat-standardized lean weight and to evaluate accuracies and precisions realized by use of equations based on variables derived from different technologies. Independent variables used to establish regression equations represented technologies of direct carcass measurements, optical probe data, TOBEC (total body electrical conductivity) readings, and dissected (DHMLN) and fat-standardized (FSHMLN) ham lean. Genotype bias existed when any equation from a single technology was used and was minimized by combining FSHMLN with one TOBEC reading, carcass length, and the probe measurement of 10th rib fat depth. Large RAC biases appeared when equations from direct carcass measurements or optical probe data were used and were minimized by an equation using either DHMLN or FSHMLN. A practical equation with relatively high R2 value and small genotype and RAC biases were developed by combining TOBEC readings with direct carcass measurements of 10th rib fat depth and warm carcass weight.

Adipose Tissue

Preparation of muscle samples for comparative electron microscopy.

The interpretation of muscle structure by scanning electron microscopy (SEM) has not been consistent among various studies. Consequently, the literature is confusing with respect to the identity of T-tubules, transverse ridges, Z-disks, and intermyofibrillar connections. The objective of this research was to evaluate the effects of different methods of sample preparation and imaging on ultrastructural details of previously identified transverse structures and intermyofibrillar connections and to verify or disprove the commonality of these structures under different viewing conditions. Scanning electron microscopy coupled with a cold stage, SEM at room temperature, and transmission electron microscopy (TEM) of thin sections were most appropriate for exposing detail of inter- and intracellular structures and for measuring sarcomere length and spacing of intermyofibrillar connections. Scanning electron microscopy of samples mounted on a cold stage, fractured, and sublimed provided excellent images of meat and muscle ultrastructure and may be used in correlative microscopy. Sarcomere length and spacing between intermyofibrillar connections were similar among most specimen preparation techniques and were affected similarly by heat treatments. Results indicate that the regularly spaced transverse structures viewed by conventional SEM and the intermyofibrillar connections viewed by low-temperature SEM are Z-disks.

Animals

Effects of ractopamine, genotype, and growth phase on finishing performance and carcass value in swine: I. Growth performance and carcass merit.

A replicated factorial experiment using 183 individually fed crossbred barrows was conducted. Pigs were fed an 18.5% CP (.95% lysine) diet with 3,594 kcal of ME/kg. The effects of five genotypes (GT): 1) Hampshire (H) x (H x Duroc [D]), 2) synthetic terminal sire line, 3) (H x D) x (Landrace [L] x [Yorkshire (Y) x D]), 4) L x (Y x D), and 5) Y x L; two levels of ractopamine (RAC) treatment: 0 and 20 ppm; and three treatment weight periods (WT): 1) 59 to 100, 2) 73 to 114, and 3) 86 to 127 kg live weight on growth performance and carcass traits were evaluated. Ractopamine increased (P less than .0001) amount (FSL) and percentage (PFSL) of carcass lean standardized to 10% fat content, loin eye area (LEA), and dressing percentage (PDRES) and reduced (P less than .01) amount (DF) and percentage (PDF) of dissected fat. Magnitudes of RAC effects were smaller than those reported by other researchers. Effects of GT and WT on all growth and carcass traits were highly significant (P less than .001) except for those of WT on ADG (P less than .05) and GT on average feed intake (AFI) and backfat thickness at the last rib (BFLR; P less than .05). Genotype 3 performed better for most economically important traits than did GT 1 and 4, suggesting that heterosis existed in GT 3, which essentially was obtained by crossing GT 1 and 4. Among the three treatment periods, WT 2 had the highest ADG. As BW increased from WT 1 to 3, AFI and AFI/ADG ratio (FCR) increased and lean percentage decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue

Effects of ractopamine, genotype, and growth phase on finishing performance and carcass value in swine: II. Estimation of lean growth rate and lean feed efficiency.

A replicated factorial experiment using 183 individually fed crossbred barrows was conducted. The pigs were fed an 18.5% CP (.95% lysine) diet with 3.594 kcal of ME/kg. The effects of five genotypes (GT): 1) Hampshire (H) X (H X Duroc [D]), 2) synthetic terminal sire line, 3) (H X D) X (Landrace [L] X [Yorkshire (Y) X D]), 4) L X (Y X D), and 5) Y X L; two levels of ractopamine (RAC) treatment: 0 and 20 ppm; and three treatment weight periods (WT): 1) 59 to 100, 2) 73 to 114, and 3) 86 to 127 kg live weight on ADG of dissected lean (ADLG) and fat standardized lean adjusted to 10% fat content (ADSLG) and feed efficiency of ADLG (LFE) and ADSLG (SLFE) were evaluated. Initial carcass lean quantity of each individual animal was determined by a regression equation (R2 = .95) generated from 30 additional barrows (six per GT) slaughtered at 59 kg and 30 (six per GT) untreated pigs slaughtered at 100 kg average live weight. Logarithmic and reciprocal transformations of dependent variables were used to stabilize heterogeneous variances and to improve normality of the residuals. Ractopamine increased (P less than .0001) ADLG, ADSLG, LFE, and SLFE, respectively, by 19.5, 25.0, 19.6, and 25.5%. Differences (P less than .001) were observed among genotypes for all traits, showing that considerable variation existed in the data and indicating that genetic improvement can be realized through the identification and selection of superior genotypes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Practical means for estimating pork carcass composition.

Three hundred sixty-one market-weight barrow and gilt carcasses were physically dissected into bone, skin, fat and muscle. A three-variable multiple linear regression equation containing the same independent variables (warm carcass weight, 10th rib loin muscle area and 10th rib fat depth) used (U.S.) to determine pork carcass lean weight was found to be the most practical means for predicting weight of muscle standardized to 10% fat. Multiple linear regression equations containing more than three independent variables produced only slight improvements in R2 values; however, the standard deviation about the regression line was not greatly improved by the addition of more independent variables to this three-independent-variable regression model. A single multiple linear regression equation using the three independent variables above may not be adequate to describe variation over the entire live-weight range for all hogs marketed in the U.S. For most accurate muscle weight prediction, different equations should be used for weight subclasses with one equation for carcasses under 100 kg and another for those heavier than 100 kg. A single prediction equation for muscle weight was adequate for carcasses of both barrows and gilts.

Adipose Tissue

Isolation of Yersinia enterocolitica and related species from porcine samples obtained from an abattoir.

Swabs of swine carcasses and samples of porcine tongue and trim obtained from an abattoir were examined for the presence of Yersinia enterocolitica and related species (Y. intermedia, Y. kristensenii and Y. frederiksenii). Three enrichment media (phosphate buffered saline, sorbitol-bile salts-phosphate buffered saline, and a modified Rappaport's broth) were compared at 4 degrees C for their efficiency of recovery of Y. enterocolitica and related species. Two secondary enrichment procedures (post-enrichment in modified Rappaport's broth for 2 d at 25 degrees C and treatment with 0.5% KOH in 0.5% NaC1) also were evaluated. The porcine isolates were characterized by biochemical and serological examination, speciation, and biotyping. Eight of 43 samples were positive for Y. enterocolitica and related species. The combination of incubation in sorbitol-bile salts-phosphate buffered saline for 21 d at 4 degrees C followed by post-enrichment in modified Rappaport's broth yielded maximum number of isolates. All isolates, except one, were avirulent as determined by autoagglutination, calcium dependence at 37 degrees C, and HeLa cell invasiveness tests.

Animals