Artifactual changes in PCV, hemoglobin concentration, and cell counts in bovine, caprine, and porcine blood stored at room and refrigerator temperatures

BACKGROUND: Blood samples collected from farm animals for hematology testing may not reach the laboratory or be examined immediately upon collection, and in some cases may need to be transported for hours before reaching a laboratory. OBJECTIVE: The objective of this study was to investigate the artifactual changes that may occur in PCV, hemoglobin (Hgb) concentration, and cell counts in bovine, caprine, and porcine blood samples stored at room (30 degrees C) or refrigerator (5 degrees C) temperature. METHODS: Baseline values for PCV, Hgb concentration, and RBC and WBC counts were determined immediately after blood collection from 36 cattle, 32 goats, and 48 pigs using manual techniques. Blood samples were split into 2 aliquots and stored at 30 degrees C or 5 degrees C. Hematologic analyses were carried out at specified intervals during 120 hours of storage. Results were analyzed by repeated measure ANOVA; results at different temperatures were compared by paired t-tests. RESULTS: Compared to baseline values, there were no significant changes in Hgb concentration, RBC count, or WBC count in samples from cattle; in Hgb concentration and RBC count in samples from goats; and in Hgb concentration and WBC count in samples from pigs throughout the 120 hours of storage at both 30 degrees C and 5 degrees C. Significant changes (P <.05) from baseline occurred in PCV after 14 hours of storage at 30 degrees C and after 19 hours of storage at 5 degrees C in cattle and goats; and after 10 hours of storage at 30 degrees C and 14 hours of storage at 5 degrees C in pigs. Significant changes also were observed in Hgb concentration at 96 hours at 30 degrees C and 5 degrees C, and in RBC counts at 48 hours at 30 degrees C and 96 hours at 5 degrees C in porcine samples; and in total WBC counts at 120 hours at 30 degrees C and 5 degrees C in caprine samples. Artifactual changes were more pronounced in the samples stored at 30 degrees C. CONCLUSIONS: At both 30 degrees C and 5 degrees C, blood samples from cattle and goats can be stored for up to 12 hours, while blood samples from pigs can be stored for up to 8 hours without any significant changes in PCV. Blood samples from all 3 species can be stored for more than 24 hours without significant changes in Hgb concentration, RBC count, and total WBC count.     

Artifactual changes in canine blood following storage, detected using the ADVIA 120 hematology analyzer

BACKGROUND: Artifactual changes in blood may occur as a consequence of delayed analysis and may complicate interpretation of CBC data. OBJECTIVE: The aim of this study was to characterize artifactual changes in canine blood, due to storage, using the ADVIA 120 hematology analyzer. METHODS: Blood samples were collected into EDTA from 5 clinically healthy dogs. Within 1 hour after blood sample collection and at 12, 24, 36 and 48 hours after storage of the samples at either 4 degrees C or room temperature (approximately 24 degrees C), a CBC was done using the ADVIA 120 and multispecies software. A linear mixed model was used to statistically evaluate significant differences in values over time, compared with initial values. RESULTS: The HCT and MCV were increased significantly after 12 hours of collection at both 4 degrees C and 24 degrees C, and continued to increase through 48 hours. The MCHC initially decreased significantly at 12-24 hours and then continued to decrease through 48 hours at both temperatures. Changes in HCT, MCV, and MCHC were greater at 24 degrees C than at 4 degrees C at all time points. A significant increase in MPV and a decrease in mean platelet component concentration were observed at all time points at 24 degrees C. Samples stored at 24 degrees C for 48 hours had significantly higher percentages of normocytic-hypochromic RBCs, and macrocytic-normochromic RBCs, and lower platelet and total WBC counts. CONCLUSIONS: Delayed analysis of canine blood samples produces artifactual changes in CBC results, mainly in RBC morphology and platelet parameters, that are readily detected using the ADVIA 120. Refrigeration of specimens, even after 24 hours of storage at room temperature, is recommended to improve the accuracy of CBC results for canine blood samples.     

Changes in gas composition and acid-base values of venous blood samples stored under different conditions in 4 domestic species

BACKGROUND: The effect of storage temperature and time on blood gas and acid-base values has been investigated intensively in cattle and dogs; however, data are lacking in other species. OBJECTIVE: The aim of our study was to evaluate changes in gas composition and acid-base values in venous blood stored at different temperatures and for different times in 4 domestic species in Italy. METHODS: Blood samples from Comisana sheep (n = 10), Maltese goats (n = 10), Ragusana donkeys (n = 10), and Thoroughbred horses (n = 10) were analyzed after storage at 23 degrees C (room temperature) for 15 minutes (group I), 23 degrees C for 1 hour (group II), 37 degrees C for 8 hours (group III), and 4 degrees C for 24 hours (group IV). Results were analyzed using a 1-way repeated measures ANOVA. RESULTS: In all species no statistically significant differences in pH values were present in samples stored at 4 degrees C for 24 hours. This also was true for PCO2 in all species except the horse. Except for HCO3- concentration in the horse, significant changes in PO2, HCO3- concentration, base excess, and the standard bicarbonate concentration were observed for all species in samples stored at 4 degrees C. In samples stored for only 1 hour at room temperature, significant changes in most analytes were detected. CONCLUSIONS: The results of this study underline the need for rapid assessment of acid-base samples, because any delay, even for 1 hour, may affect the results.     

Effects of temperature and handling conditions on lipid emulsion stability in veterinary parenteral nutrition admixtures during simulated intravenous administration

OBJECTIVE: To determine whether lipid particle coalescence develops in veterinary parenteral nutrition (PN) admixture preparations that are kept at room temperature (23 degrees C) for > 48 hours and whether that coalescence is prevented by admixture filtration, refrigeration, or agitation. SAMPLE POPULATION: 15 bags of veterinary PN solutions. PROCEDURES: Bags of a PN admixture preparation containing a lipid emulsion were suspended and maintained under different experimental conditions (3 bags/group) for 96 hours while admixtures were dispensed to simulate IV fluid administration (rate, 16 mL/h). Bags were kept static at 4 degrees C (refrigeration); kept at 23 degrees C (room temperature) and continuously agitated; kept at room temperature and agitated for 5 minutes every 4 hours; kept static at room temperature and filtered during delivery; or kept static at room temperature (control conditions). Admixture samples were collected at 0, 24, 48, 72, and 96 hours and examined via transmission electron microscopy to determine lipid particle diameters. At 96 hours, 2 samples were collected at a location distal to the filter from each bag in that group for bacterial culture. RESULTS: Distribution of lipid particle size in the control preparations and experimentally treated preparations did not differ significantly. A visible oil layer developed in continuously agitated preparations by 72 hours. Bacterial cultures of filtered samples yielded no growth. CONCLUSIONS AND CLINICAL RELEVANCE: Data indicated that the veterinary PN admixtures kept static at 23 degrees C are suitable for use for at least 48 hours. Manipulations of PN admixtures appear unnecessary to prolong lipid particle stability, and continuous agitation may hasten lipid breakdown.     

The effectiveness of gentamicin or polymyxin B for the control of bacterial growth in equine semen stored at 20 degrees C or 5 degrees C for up to forty-eight hours.

Semen from three stallions was used to evaluate the effectiveness of two antibiotics added to semen extender for samples stored at 20 degrees C or 5 degrees C for up to 48 hours. Each ejaculate was divided into six different treatments: semen+extender (SE); SE+gentamicin (100 micrograms/mL); SE+polymyxin B (1000 units/mL); and each of the above treatments inoculated with Pseudomonas aeruginosa ATCC 27853. Sampling of diluted semen for bacteriological analysis was performed after 2, 8, 24 and 48 hours of preservation at either temperatures. The presence of nonspecific bacteria was noted after two hours in all SE aliquots. The number of bacteria did not change in samples stored at 5 degrees C, while in samples preserved at 20 degrees C, it increased by three to four times after 48 hours. In semen aliquots treated with either of the antibiotics, the number of nonspecific bacteria was very low after two and eight hours at both temperatures. This number remained stable up to 48 hours at 5 degrees C, while an increase was noted at 24 and 48 hours at 20 degrees C. At 5 degrees C, the number of P. aeruginosa cells tended to decrease between 24 and 48 hours in SE aliquots. The presence of gentamicin or polymyxin B appeared to rapidly inhibit growth of P. aeruginosa. At 20 degrees C, growth of P. aeruginosa increased between 8 and 24 hours in SE, while the presence of antibiotics almost completely inhibited the growth of the bacterium.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of an electrolyte analyzer for measurement of ionized calcium and magnesium concentrations in blood, plasma, and serum of dogs

OBJECTIVE: To evaluate an electrolyte analyzer for measurement of ionized calcium (Ca(i)) and magnesium (Mg(i)) concentrations in blood, plasma, and serum; investigate the effect of various factors on measured values; and establish reference ranges for Ca(i) and Mg(i) in dogs. ANIMALS: 30 healthy adult dogs of various breeds. PROCEDURE: Precision in a measurement series, day-to-day precision, and linearity were used to evaluate the analyzer. The effects of exposure of serum samples to air, type of specimen (blood, plasma, or serum), and storage temperature on sample stability were assessed. Reference ranges were established with anaerobically handled serum. RESULTS: The coefficient of variation for precision in a measurement series was < or = 1.5% for both electrolytes at various concentrations. The Ca(i) and Mg(i) concentrations were significantly lower in aerobically handled serum samples, compared with anaerobically handled samples. The Ca(i) and Mg(i) concentrations differed significantly among blood, plasma, and serum samples. In anaerobically handled serum, Ca(i) was stable for 24 hours at 22 degrees C, 48 hours at 4 degrees C, and 11 weeks at -20 degrees C; Mg(i) was stable for 8 hours at 22 degrees C, < 24 hours at 4 degrees C, and < 1 week at -20 degrees C. In anaerobically handled serum, reference ranges were 1.20 to 1.35 mmol/L for Ca(i) and 0.42 to 0.58 mmol/L for Mg(i). CONCLUSIONS AND CLINICAL RELEVANCE: The electrolyte analyzer was suitable for determination of Ca(i) and Mg(i) concentrations in dogs. Accurate results were obtained in anaerobically handled serum samples analyzed within 8 hours and kept at 22 degrees C.     

Effects of temperature and duration of sample storage on the haematological characteristics of western grey kangaroos (Macropus fuliginosus)

OBJECTIVE: To investigate the effects of storage duration and temperature on haematological analyses performed on blood from the western grey kangaroo (Macropus fuliginosis). METHOD: Blood samples from five western grey kangaroos were stored at 4 degrees C, 24 degrees C and 36 degrees C. Each sample was analysed haematologically over a 5-day period. RESULTS: The blood samples maintained optimal stability at 4 degrees C. At this temperature the haematological values remained essentially unchanged for the duration of the study, while samples stored at 36 degrees C and 24 degrees C showed significant changes in some haematological measures by 12 h and 48 h, respectively. Disturbances in leukocyte morphology were evident, to varying degrees, in all samples. CONCLUSIONS: Blood samples from macropodids should be tested within 48 h of collection if stored at a room temperature of about 24 degrees C. Where testing is to be delayed for more than 48 h, samples should be refrigerated as soon as possible. Exposure of samples to heat in excess of 24 degrees C should be avoided at all times.     

Effects of season and sample handling on measurement of plasma alpha-melanocyte-stimulating hormone concentrations in horses and ponies

OBJECTIVE: To investigate effects of sample handling, storage, and collection time and season on plasma alpha-melanocyte-stimulating hormone (alpha-MSH) concentration in healthy equids. ANIMALS: 11 healthy Standardbreds and 13 healthy semiferal ponies. PROCEDURE: Plasma alpha-MSH concentration was measured by use of radioimmunoassay. Effects of delayed processing were accessed by comparing alpha-MSH concentrations in plasma immediately separated with that of plasma obtained from blood samples that were stored at 4 degrees C for 8 or 48 hours before plasma was separated. Effects of suboptimal handling were accessed by comparing alpha-MSH concentrations in plasma immediately stored at -80 degrees C with plasma that was stored at 25 degrees C for 24 hours, 4 degrees C for 48 hours or 7 days, and -20 degrees C for 30 days prior to freezing at -80 degrees C. Plasma alpha-MSH concentrations were compared among blood samples collected at 8:00 AM, 12 noon, and 4:00 PM. Plasma alpha-MSH concentrations were compared among blood samples collected in January, March, April, June, September, and November from horses and in September and May from ponies. RESULTS: Storage of blood samples at 4 degrees C for 48 hours before plasma was separated and storage of plasma samples at 4 degrees C for 7 days prior to freezing at -80 degrees C resulted in significant decreases in plasma alpha-MSH concentrations. A significantly greater plasma alpha-MSH concentration was found in September in ponies (11-fold) and horses (2-fold), compared with plasma alpha-MSH concentrations in spring. CONCLUSIONS AND CLINICAL RELEVANCE: Handling and storage conditions minimally affected plasma alpha-MSH concentrations. Seasonal variation in plasma alpha-MSH concentrations must be considered when evaluating pituitary pars intermedia dysfunction in equids.     

Stability of sorbitol dehydrogenase activity in bovine and equine sera

Serum sorbitol dehydrogenase (SDH) activities in 10 cows and nine horses were measured using an automated clinical analyzer. The serum samples were divided into aliquots that were stored at room temperature (21 degrees C), refrigerated (0-5 degrees C), or frozen (-30 degrees C). The stability of the SDH activity was monitored at various intervals. SDH activity in bovine sera remained stable for at least 5 hours at room temperature, 24 hours refrigerated, and 72 hours frozen without any significant (p < 0.05) differences from the initial serum values. In equine sera, SDH activity remained stable for at least 5 hours at room temperature and 48 hours frozen. The activity of the refrigerated equine sera was stable for at least 5 hours but less than 24 hours. An evaluation of fresh bovine serum and heparinized plasma samples indicated that there was no significant difference (p < 0.05) between the two sampling methods and that either may be employed for automated measurement of SDH activity following the established protocol. Sample type comparison indicated that there was a small but statistically significant (p < 0.05) difference between the results obtained comparing fresh serum and heparinized plasma samples for the horse. A reference range for Holstein cows was established using sera from 71 clinically healthy cattle (mean -/+ 2 SD = 32 -/+ 26 U/L).     

Changes in the acid-base equilibrium in cattle in relation to the time intervals between sampling and study under field conditions

The parameters of acid-base balance were investigated in relation to the time difference between the blood sampling and the examination and ambient temperature in the field conditions. The transfer of the samples from field conditions to a laboratory was imitated by putting the case with the samples into a thermostat at the temperature of 20 degrees C and 37 degrees C. In the first trial, at the temperature of 20 degrees C, statistically significant changes in pH values were recorded in seven hours. If the temperature in the case was 37 degrees C, the differences in pH, BE, SBi and BB values and in the partial pressure of CO2 were statistically significant in six hours. Applying the above results we state that the acid-base parameters can be examined within five hours after blood sampling supposing that the blood samples are kept at the temperature of 0-4 degrees C and using for veterinary diagnostics the equilibration method after Astrup.     

The effect of thermal environment and age on neonatal pig behavior

Three experiments were conducted to evaluate the ability of a radiant environment and the presence of a littermate to attract pigs during the first 3 d of age. The effect of stimuli on pig movement was studied in an enclosed rectangular aluminum test chamber containing four similar sections that were heated independently. In Exp. 1, all sections were at 34.8 degrees C to evaluate the chamber for biases of where pigs located themselves at 1 (n = 24) and 2 d (n = 26) of age. More (P < .025) pigs settled (e.g., no movement for 7 min) in end sections than in middle sections. Age did not affect time to settle or settling location. The effect on pig location of heating one chamber end section to either 23, 40, 48, 56, or 64 degrees C and leaving the remaining sections unheated (24 degrees C) was determined in Exp. 2. Settling of pigs at 1 (n = 50) and 2 d (n = 50) of age was affected by temperature (P < .001) but not by age. The minimum distance between average pig location and the heated section occurred at 48 degrees C. Experiment 3 involved 15 pigs each at 1 and 3 d during a 1-h trial to compare the relative pig attraction to 1) a heated chamber end section at 44.4 degrees C when remaining sections were at 23.5 degrees C, 2) an anesthetized littermate in an end section when all sections were at 24.1 degrees C, or 3) a choice test involving a 45.5 degrees C end section and an anesthetized littermate in the opposite end section with three unheated sections at 23.7 degrees C. Average distance between the test animal and the heated section was greater (P < .01) than that between the test animal and an anesthetized pig. Pigs that were allowed a choice preferred to lie near an anesthetized littermate in a cold section rather than alone in a 45 degrees C section (P < .01), and they were less (P < .005) active when an anesthetized littermate was present in the chamber. Although radiant heat effectively attracted pigs, heat was less attractive than an anesthetized littermate. The greater attraction of pigs to a littermate than to radiant heat may explain why pigs remain near the sow and littermates during d 1 and 2 after birth.     

The effect of storage temperature on the second day methylene-blue test.

The purpose was to show whether or not the methylene-blue test can be postponed to the second day. The milk samples were preserved at three different temperatures 8-10 degrees C, 5-7 degrees C and 2-4 degrees C and the storage time was 16-22 hours. The test indicated that the methylene-blue test could be postponed to the next day satisfying the practical needs if the samples were kept at temperature of 2-4 degrees C. At this temperature the results of the second day methylene-blue test differed statistically not more than p less than 0.05 (nearly significant) from the results of the first day methylene-blue test and the percentage of quality class changes was 2.9.     

Use of the tetrazolium salt reduction test in the detection of phagocytic activity in pigs

One of the indicators of non-specific, cell-mediated immunity can be the ability of phagocytic system to react to the presence of antigenic impulses and stimulators. The phagocytic activity induced in this way in vitro can be evaluated quantitatively by help of different methods. In this study the method of the reduction of colourless tetrazolium salt (INT) to the red formazane was used. The optimum conditions for carrying out this test in isolated peripheral pig leucocytes were determined. It was stated that 3-4 ml samples of pig blood, from which 6.10(6) leucocytes necessary for the test can be isolated, were sufficient for the examination of phagocytic activity. Further, 45 minutes were determined as the optimum time of incubation It was found that the INT test can be reproduced by 24 hours after blood sampling provided the blood is kept at the temperature of +4 degrees C. The INT test was used for examining 38 pig blood samples before and after myostress; this stress has a significant effect both on a decrease of phagocytic activity and on the ability of leucocytes to migrate in the LMI test. The INT test was further used for examining pigs before the administration of the first dose of the inactivated vaccine against the Aujeszky's disease virus and two days after; the application of the vaccine significantly increased the phagocytic activity.     

Stability of hematologic analytes in monkey, rabbit, rat, and mouse blood stored at 4°C in EDTA using the ADVIA 120 hematology analyzer

Background: The time from sampling to analysis can be delayed when blood samples are shipped to distant reference laboratories or when analysis cannot be readily performed. Objective: The objective of this study was to evaluate the stability of hematologic analytes in blood samples from monkeys, rabbits, rats, and mice when samples were stored for up to 72 hours at 4°C. Methods: Blood samples from 30 monkeys, 15 rabbits, 20 rats, and 30 mice were collected into EDTA‐containing tubes and were initially analyzed within 1 hour of collection using the ADVIA 120 analyzer. The samples were then stored at 4°C and reanalyzed at 24, 48, and 72 hours after collection. Results: Significant (P<.0003) changes in hematologic analytes and calculations included increased HCT and MCV and decreased MCHC and cell hemoglobin concentration mean (CHCM) at 72 hours and increased MPV at 24 hours in monkeys; increased MCV at 72 hours and MPV at 48 hours and decreased monocyte count at 24 hours in rabbits; increased MCV and decreased MCHC, CHCM, and monocyte count at 24 hours in rats; increased MCV, red cell distribution width, and MPV and decreased MCHC, CHCM, and monocyte count at 24 hours in mice. Conclusions: Although most of the changes in the hematologic analytes in blood from monkeys, rabbits, rats, and mice when samples were stored at 4°C were analytically acceptable and clinically negligible, the best practice in measuring hematologic analytes in these animals is timely processing of blood samples, preferably within 1 hour after collection.     

Observations on the cooling and cryopreservation of pig oocytes at the germinal vesicle stage

This study examined the viability of pig oocytes at the germinal vesicle stage following cooling or cryopreservation. Cumulus-intact oocytes (n = 641) were collected from slaughterhouse pig ovaries and used in two experiments. In Exp. I the viability of 1) control, 2) cryoprotectant control (CC, 1.5 M glycerol/.5 M sucrose), 3) cooled (0 degrees C) and 4) cryopreserved (-196 degrees C) oocytes was assessed after no incubation or a 24-h incubation. Survivability was judged by morphological appearance, trypan blue exclusion and fluorescein diacetate staining. Survival rate of control oocytes (90%; based primarily on morphological appearance of the cumulus) incubated 0 h was greater (P less than .05) than that of all other groups, whereas survival rate of -196 degrees C oocytes (57%) was less (P less than .05) than that of all other groups. However, vital staining of 0 degrees C and -196 degrees C oocytes showed 0% survival rate as evidenced by trypan blue uptake and lack of fluorescence. The cumulus cells surrounding oocytes that were stored at 0 degrees C or -196 degrees C survived freezing as evidenced by trypan blue exclusion and intense fluorescence. Similar differences among treatment groups were found for oocytes incubated 24 h. Exp. 2 examined the temperature at which oocytes became sensitive to cooling. Oocyte death occurred when oocytes were cooled to 15 degrees C or lower. These results demonstrate that pig oocytes at the germinal vesicle stage did not survive cooling to 15 degrees C or below. When assessing the viability of cryopreserved cumulus enclosed oocytes it is important to use vital stains in conjunction with morphological appearance.     

Measurement of prothrombin time (PT) and activated partial thromboplastin time (APTT) on canine citrated plasma samples following different storage conditions

Samples from 75 clinically ill dogs were utilised in the study. APTT and PT tests were performed immediately on fresh citrated plasma samples (Fresh). The remaining plasma was stored at -20 degrees C for less than 4 months (n=36 samples) or between 4 and 7 months (n=39 samples). In batches of five, frozen samples were thawed rapidly and APTT and PT tests were performed on the thawed samples immediately (0RT) and after storage at room temperature (23 degrees C, range: 22-25 degrees C) for 24h (24RT) and 48h (48RT). The median APTT value from the (0RT) samples was significantly longer than that obtained from fresh samples (15s vs. 13.2s) but the PT value was not statistically different (7.8s vs. 7.6s). The median APTT (15s) and PT (7.5s) results from the (24RT) samples were not statistically different to those from the (0RT) samples (APTT: 15s, PT: 7.6s) but both tests were significantly longer (APTT: 16.5s, PT: 9.2s) from the (48RT) samples. We concluded that long term batching and freezing of clinical samples at -20 degrees C is acceptable for measurement of PT but not of APTT. We demonstrated that APTT and PT results do not change following storage of samples at room temperature for 24h but storage for 48h may lead to statistically and clinically significant changes (values at least 25% higher than the high value of the laboratory's reference interval) in both clotting times.     

Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.

Although Clostridium difficile is recognized as a cause of enterocolitis in horses and humans, there has been little work published regarding the lability of C. difficile and its toxins in feces. A significant decrease in recovery of C. difficile from inoculated equine fecal samples occurred during storage. Recovery after storage in air at 4 degrees C decreased from 76% (37/49) after 24 hours to 67% (33/49) at 48 hours and 29% (14/ 49) after 72 hours. In contrast to aerobic storage, 25 of 26 samples stored anaerobically at 4 degrees C yielded growth of C. difficile for 30 days, whereas the organism was only detected for 2.5 +/- 2.52 days (x +/- SD) in paired samples stored aerobically. The use of an anaerobic transport medium was effective in maintaining viability of C. difficile. These findings indicate that poor aerotolerance is the reason for the rapid decrease in culture yield. In contrast to C. difficile organisms stored aerobically at 4 degrees C, C. difficile toxins were considerably more stable and could be detected by enzyme-linked immunosorbent assay in both broth and inoculated fecal samples for at least 30 days. The poor survival of C. difficile but the stability of its toxins when feces are stored aerobically must be considered when submitting samples for diagnosis of C. difficile-associated enterocolitis in horses and when interpreting laboratory results.     

Comparison of postmortal changes in the ultrastructure of the masseter muscle and the longissimus dorsi muscle in pigs with PSE-meat]

Samples of the two muscles were taken from 8 Landrace fattened pigs, affected with pale, soft exudative meat, during stunning and 15, 30, 45 and 60 minutes after death; also 2, 3, 6, 12, 24 and 48 hours after slaughter, with storage at 20 degrees C for the first six hours and 2-5 degrees C subsequently. Considerable changes were found during the first hour after death, or even in samples taken during stunning, in longissimus dorsi samples. These consisted of destruction of mitochondria and sarcoplasmic reticulum, breakdown of cell membranes, liberation of clumps of fibre protein, disappearance of glycogen, rigor and destruction of capillaries. Such changes would account for the features of pale, soft exudative meat such as water loss, brief rigor, pale colour, deficiency of energy-rich phosphates. By contrast, in the masseter muscles of the same animal these changes did not occur until later, or were in port absent. In both muscle the breakdown of fibres took place by destruction of the "I" bands and the "Z" strips, and this process also commenced in longissimus dorsi before masseter.     

Temporal effects of 3 commonly used anticoagulants on hematologic and biochemical variables in blood samples from macaws and Burmese pythons

BACKGROUND: Few studies have been done to evaluate anticoagulants for use with blood samples from birds and reptiles. Heparin currently is the most commonly used anticoagulant in practice, but may adversely affect blood cell staining and quantitation. OBJECTIVE: The purpose of this study was to evaluate the effects of lithium heparin, K3-EDTA, and sodium citrate, with and without the addition of albumin, on hematologic variables in macaw (Ara sp) and python (Python molurus bivittatus) blood samples. METHODS: Blood samples from 10 macaws and 10 Burmese pythons were collected in heparin-coated syringes and placed into tubes containing either lithium heparin, K3-EDTA, or sodium citrate with and without the addition of 0.25 mL of a 22% bovine serum albumin solution. Cell lysis was determined by counting the number of lysed cells/200 WBCs in Wright's-Giemsa-stained blood smears and by qualitative evaluation of pink plasma in microhematocrit tubes. A CBC was done after 3, 12, and 24 hours of storage at 4 degrees C in anticoagulant-containing tubes and results were compared with those obtained at 0 hour for the heparin-coated syringe sample. A biochemical panel also was done at each time point in similarly stored lithium-heparin samples. RESULTS: Hemolysis was significantly increased in citrated samples from both macaws and pythons beginning at 12 hours. At 24 hours, 19 of 30 (63%) macaw samples in all anticoagulants had >100 lysed cells/200 WBCs. There were no significant differences in hematologic values in samples from pythons collected in heparin or EDTA at any time point. No significant differences were found in the number of lysed cells or in other hematologic data in samples with albumin. Glucose concentration decreased and potassium concentration increased significantly over time in heparinized blood samples. CONCLUSIONS: Based on the results of this study, whole blood samples anticoagulated with lithium heparin or EDTA should be evaluated within 12 hours (macaws) or 24 hours (pythons) of collection and stored at 4 degrees C for best results. Citrate should be avoided as it may result in increased cell lysis. The addition of albumin does not prevent cell lysis.     

Tests of Milkofix, a new preservative preparation for milk samples used for infrared analysis of milk components. I. Verification of its bacteriostatic and bacteriocidal effects and its interference effects]

Hygienic, ecological and health problems of sample preservation for an analysis of basic milk components make us continually to develop a safer chemical preservative substance which will preserve the original sample composition for the time required and which will not influence the analyses. Trzicky (1990) proposed Milkofix (M), a preservative substance on the basis of silver compound. The author reports on minimum risks of the use of this preparation, in comparison with traditional preservatives. Preservative efficiency of Milkofix was compared with other preservatives: K2Cr2O7 (C), NaN3 (A) and bronopol (B). The following concentrations were used: A--0.0085 g NaN3 and 0.0630 g NaCl, B--0.0050 g bronopol and 0.0500 g NaCl, C--0.0330 g K2Cr2O7 and 0.0670 g KCl in tablet, M--0.1250 g of the mixture, all amounts are per 25 ml milk. The observed antibacterial efficiency of M could be seen in a slower decrease in actual acidity, and/or in an increase in titratable acidity in M-treated samples unlike untreated ones (N). From the starting value pH 6.3 (Fig. 1), the value of N treatment dropped to 3.8 after two days, the values of M and A treatments dropped to 4.9 after nine days and to 5.7 after twelve days, respectively. As for SH, the values increased within the same interval from 6.5 (2.5 mmol/l) to 28.6 in N, and to 22.3 in M and 9.4 in A (Fig. 3). There was a similar trend when the milk samples were stored at a temperature of 4 degrees C, but the differences between the preservation methods were not so clear in comparison with storage at a temperature of 20 degrees C (Figs. 1 and 3). The standardized SH value of 9.0 (2.5 mmol) for infraanalyzer measurements was exceeded after 24 hours in N samples, after four days in M samples and after 12 days in A samples at a temperature of 20 degrees C. The observation of the growth of microorganism counts (CPM) showed that this growth was slower in M than in N, but faster in the samples of C treatment (Fig. 5). The generative time of CPM in N made 1.6 hours, in M 2.4 hours and in C 7.9 hours. The lag phase of these mixed cultures was 24 hours in M, 60 hours in C and in N treatment the lag phase was zero.(ABSTRACT TRUNCATED AT 400 WORDS)