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PubMed1990

Hematopoietic growth factors: overview and clinical applications, Part II.

The growth and differentiation of blood cells is regulated by a group of at least 12 glycoproteins, collectively known as hematopoietic growth factors. Advances in protein biochemistry and molecular genetics have provided the tools for the bulk production of these hormones for clinical application. Clinical trials of macrophage colony-stimulating factor, granulocyte macrophage colony-stimulating factor, granulocyte colony-stimulating factor, and interleukin-3 have all demonstrated significant effects on the peripheral blood counts of the recipients. The clinical usefulness of at least two of these agents in ameliorating post-chemotherapy myelosuppression, in the treatment of other cytopenias, and in enhancing engraftment after bone marrow transplantation has already been demonstrated. Potential applications to the therapy and diagnosis of other clinical disorders is under study. The history of the elucidation of these growth factors, our current understanding of their properties, interactions, and clinical effects, and the potential prospects for their future use in the manipulation of human blood cell production are the subject of this review.

Colony-Stimulating Factors↗

PubMed1990

Hematopoietic growth factors: overview and clinical applications, Part III.

Animals↗

PubMed1993

Effects of recombinant human interleukin-11 on hematopoietic reconstitution in transplant mice: acceleration of recovery of peripheral blood neutrophils and platelets.

We have examined the effects of recombinant human interleukin-11 (rhIL-11) on the recovery of peripheral blood cell counts and proliferation of progenitors and hematopoietic stem cells (day 12 colony-forming units-spleen-CFU-S12) in vivo using a mouse bone marrow (BM) and spleen cell transplantation model. Recovery of leukocytes was accelerated in animals receiving daily administration of rhIL-11 (100 micrograms/kg/d) and reached normal levels by day 14 posttransplantation. This increased total leukocyte count reflected mainly an increase in neutrophils. Neutropenia (absolute neutrophil count [ANC] < 1,500) was present in control transplant mice for 14 to 15 days, while in the rhIL-11-treated group, neutrophils recovered to normal by days 8 to 10 and continued to increase until day 19. Animals treated with rhIL-11 had only 1 day with ANC demonstrated < 500. Correspondingly, rhIL-11 treatment increased granulocyte-macrophage progenitors (CFU-GM) derived from both spleen and BM cells. Higher doses of IL-11 increased CFU-GM nearly threefold and CFU-Mix fourfold to fivefold, while increasing burst-forming units-erythroid to a lesser degree. BM and spleen cellularity were both increased in IL-11-treated mice, but no increase in CFU-S12 was noted. In addition, in vivo daily administration of IL-11 increased peripheral platelet counts by threefold over control transplant mice at day 10 posttransplantation during the post-irradiation platelet nadir. Further treatment led to platelet counts higher than normal 18 days posttransplantation when control animals had just attained normal platelet counts. IL-11 can accelerate the recovery of the peripheral blood leukocytes, mainly neutrophils, and platelets in transplant mice, effects that may be clinically useful in future applications for BM transplantation and chemotherapy-related cytopenias.

Animals↗

PubMed1994

Automated reticulocyte analysis. Clinical practice and associated new parameters.

Automated reticulocyte analysis, through continued instrument, software, and reagent developments, provides improved precision for the reticulocyte count. This rapid and simple testing assists in providing a cost effective and sensitive approach to the diagnosis and therapeutic monitoring of the anemic patient. As laboratory and clinical hematologist become familiar with the benefits and expanding clinical applications of this technology, the use of reticulocyte analysis will increase.

Autoanalysis↗

PubMed2003

Measuring hospital quality: can medicare data substitute for all-payer data?

OBJECTIVES: To assess whether adverse outcomes in Medicare patients can be used as a surrogate for measures from all patients in quality-of-care research using administrative datasets. DATA SOURCES: Patient discharge abstracts from state data systems for 799 hospitals in 11 states. National MedPAR discharge data for Medicare patients from 3,357 hospitals. State hospital staffing surveys or financial reports. American Hospital Association Annual Survey. STUDY DESIGN: We calculate rates for 10 adverse patient outcomes, examine the correlation between all-patient and Medicare rates, and conduct negative binomial regressions of counts of adverse outcomes on expected counts, hospital nurse staffing, and other variables to compare results using all-patient and Medicare patient data. DATA COLLECTION/EXTRACTION: Coding rules were established for eight adverse outcomes applicable to medical and surgical patients plus two outcomes applicable only to surgical patients. The presence of these outcomes was coded for 3 samples: all patients in the 11-state sample, Medicare patients in the 11-state sample, and Medicare patients in the national Medicare MedPAR sample. Logistic regression models were used to construct estimates of expected counts of the outcomes for each hospital. Variables for teaching, metropolitan status, and bed size were obtained from the AHA Annual Survey. PRINCIPAL FINDINGS: For medical patients, Medicare rates were consistently higher than all-patient rates, but the two were highly correlated. Results from regression analysis were consistent across the 11-state all-patient, 11-state Medicare, and national Medicare samples. For surgery patients, Medicare rates were generally higher than all-patient rates, but correlations of Medicare and all-patient rates were lower, and regression results less consistent. CONCLUSIONS: Analyses of quality of care for medical patients using Medicare-only and all-patient data are likely to have similar findings. Measures applied to surgery patients must be used with more caution, as those tested only in Medicare patients may not provide results comparable to those from all-patient samples or across different samples of Medicare patients.

Aged↗

PubMed1999

Platelet aggregation during cardiopulmonary bypass evaluated by a laser light-scattering method.

BACKGROUND: In regard to postoperative bleeding, the most important consequence of cardiopulmonary bypass (CPB) is the loss of aggregability. However, the mechanism of platelet aggregation loss during CPB is unclear. Newly developed particle-counting methods that use light scattering can be used to quantify changes in the number of platelet aggregates of different sizes after application of an aggregating stimulus. Using a light-scattering method, we investigated changes in platelet aggregation during cardiac operation. METHODS: Nineteen patients undergoing CPB were evaluated. Blood samples were obtained before the operation, 1 hour after initiation of CPB, at the end of CPB, at the end of the operation, and on day 1 after the operation. Platelet aggregation after stimulation by 2.5 micromol/L adenosine diphosphate and 2.0 microg/mL collagen was determined; small (9 to 25 microm), medium (25 to 50 microm), and large (50 to 70 microm) aggregates were counted. RESULTS: Generation of medium and large aggregates after stimulation with adenosine diphosphate and collagen were significantly decreased with CPB, whereas, in spite of hemodilution, the quantity of the small aggregates was maintained at the elevated level. CONCLUSIONS: These results reflect the fact that CPB does not affect the first phase of aggregation. It suggests that platelet dysfunction associated with CPB is mainly caused by an inhibition in the development of small aggregates into larger aggregates.

Adult↗

PubMed1997

[External assessment of analytic quality in hematology: a necessity in Latin America].

The assurance of analytical quality in a clinical laboratory is achieved through an internal system of quality control complemented by an external evaluation program. Quality assurance provides a foundation for the confidence that is placed in laboratory results and their use in the diagnosis and treatment of diseases. Many laboratories in Latin American countries do not have appropriate systems in place to evaluate and control quality. Given the importance of diagnoses based on hematologic data, the Pan American Health Organization sponsored a course in quality control in hematology during the XI Latin American Congress of Clinical Biochemistry (Mexico, 1993), in which representatives from Argentina, Chile, Cuba, Mexico, Paraguay, Dominican Republic, and Uruguay participated. As part of the course, the following control materials were produced: secondary standard solution of cyanmethemoglobin, stabilized concentrated hemoglobin solution, and preserved human whole blood with pseudoleukocytes. These materials were sent to laboratories in the seven participating countries for use in analytical procedures, and the results were then subjected to an external evaluation to assess individual performance and the comparability of results among the group. The specific tasks carried out were: (1) determination of values for hemoglobin, hematocrit, and red and white blood cell counts by the procedures normally used in each laboratory; (2) recording of the data on special reporting forms; and (3) transmittal of those forms to the coordinator in each country. The results were analyzed with regard to both the procedure used and the participating country. Reference values were established by consensus following application of a statistical method to eliminate outlying values. Comparative analysis of the results showed the coefficients of variation (CV) of the hematocrit (4.5%), red blood cell count (11.0%), and white blood cell count (22.2%) to be higher than those reported from the United States of America and Europe. With regard to analytical procedures, the manual methods yielded larger CV than the automated methods. When analysis of variance (ANOVA) was used on data broken down by country and by procedure, the only statistically significant result was for leukocyte count (P < 0.02). It was concluded that training in the preparation of quality control materials and the subsequent use of these materials in pilot surveys could provide a starting point for establishing continuous internal and external quality assessment systems in hematology. Such systems, together with continuing education for laboratory personnel and the availability of automated instrumentation, will lead to achievement of optimum laboratory quality.

Hematology↗

PubMed2001

Saratin, an inhibitor of von Willebrand factor-dependent platelet adhesion, decreases platelet aggregation and intimal hyperplasia in a rat carotid endarterectomy model.

PURPOSE: Post-carotid endarterectomy, thrombosis, and intimal hyperplasia may be decreased by the inhibition of platelet adhesion and activation. In this study, a novel agent, saratin, was used to inhibit platelet-to-collagen adhesion in a rat carotid endarterectomy model. Saratin is a recombinant protein isolated from the saliva of the medicinal leech Hirudo medicinalis, which is thought to act by binding to collagen, and inhibits von Willebrand factor-collagen interaction under conditions of increased shear and therefore, the adherence and activation of platelets at the vessel wall. Saratin has the advantage of being a nonsystemic, site-specific topical application. METHODS: A rat carotid endarterectomy model was used in which an open technique with arteriotomy and intimectomy was used. Saratin was applied to the endarterectomized surface of the carotid artery before arterial closure. End point measurements included platelet adhesion, thrombosis rate, intimal hyperplasia development, bleeding times, and platelet counts. Electron micrographs of carotid arteries were used for quantitative analysis of platelet aggregation and platelet counts. Intimal hyperplasia and thrombosis were assessed with computer-assisted morphometric analysis of elastin-stained carotid artery sections with direct measurement of the intimal hyperplasia area. RESULTS: The topical application of saratin significantly decreased platelet adhesion compared with controls at 3 hours after carotid endarterectomy (64 +/- 17 vs 155 +/- 33 platelets per grid, P = .05), and 24 hours after carotid endarterectomy (35 +/- 11 vs 149 +/- 37 platelets per grid, P = .0110), respectively. A percent luminal stenosis, as a measure of intimal hyperplasia, was significantly decreased with saratin application compared with controls (10.9% +/- 1.8% vs 29.8% +/- 6.8%, P = .0042). This decrease in intimal hyperplasia formation correlated with the inhibition of platelet adhesion. Thirty-three percent of control arteries were found to be thrombosed 2 weeks after carotid endarterectomy compared with a 0% thrombosis rate in the saratin-treated group (P = .0156). No increased bleeding was encountered along the arterial suture line in the saratin group. Bleeding times and systemic platelet counts were not found to change significantly in the saratin-treated rats compared with control rats at 3 and 24 hours after endarterectomy. CONCLUSION: Saratin significantly decreased platelet adhesion, intimal hyperplasia, luminal stenosis, and thrombosis after carotid endarterectomy in rats. Saratin did not increase suture line bleeding or bleeding times, and did not decrease platelet counts. Saratin may serve as a topical agent to be used for the site-specific inhibition of thrombosis and intimal hyperplasia after vascular manipulation.

Administration, Topical↗

PubMed1992

An optical method for the determination of platelet count in platelet samples contaminated with red blood cells.

We present a simple photometric method to determine the total concentration of platelets present in a sample independently of red blood cell concentration. Standard optical density curves for platelet samples ranging in concentration from 0 to 1.5 x 10(9) cells/ml and contaminated with red blood cells ranging in concentration from 0 to 0.03 x 10(9) cells/ml are determined. A study of the absorbance spectra of red blood cells and platelets suggests that by calculating the absorbance difference between two wavelengths, an estimate of red blood cell concentration can be made. Then, in the second step of this two-step method, the individual absorbance measurements at the two wavelengths are matched to the standard values determined previously to derive an estimate of platelet concentration. In a trial of 62 unknown platelet samples contaminated with red blood cells, the standard deviation for the error in platelet count was 0.16 x 10(9) cells/ml with a mean difference of 0.011 x 10(9) platelets/ml. We conclude that our method may be useful in laboratories not equipped with electronic cell counters as well as in applications such as the development of noninvasive measurements of platelet concentration in platelet transfusion packs.

Erythrocytes↗

PubMed1992

Effect of stem cell factor with and without granulocyte colony-stimulating factor on neonatal hematopoiesis: in vivo induction of newborn myelopoiesis and reduction of mortality during experimental group B streptococcal sepsis.

Neonatal hematopoiesis and host defense are developmentally immature and under states of increased demand predispose the newborn to peripheral cytopenias and depletion of bone marrow storage pool reserves. We have previously demonstrated that recombinant human granulocyte colony-stimulating factor (rhG-CSF) can significantly modulate neonatal rat granulopoiesis and act synergistically with antibiotic therapy to reduce the mortality rate during experimental group B streptococcal sepsis. Stem cell factor (SCF) has been shown to stimulate early hematopoietic progenitor cells and, in the presence of lineage-specific CSFs, enhance committed progenitor cell proliferation. In the present study we examined the in vivo neonatal hematologic effects of recombinant rat (rr) SCF (14 days), simultaneous rrSCF + rhG-CSF (14 days), and sequential combination of rrSCF (7 days) + rhG-CSF (7 days). Sprague-Dawley newborn rats (less than or equal to 24 hours) were injected intraperitoneal (IP) x 14 days with the above combinations. rrSCF (0 to 200 micrograms/kg/d) had a negligible effect on the peripheral platelet count and absolute neutrophil count (ANC) but the diminution in the hematocrit during the first 10 days of treatment was less pronounced (P = .0001). However, the simultaneous use of rrSCF + rhG-CSF synergistically increased the circulating day 6 to 13 ANC (P = .001). Similarly, sequential rrSCF + rhG-SCF also had a synergistic significant effect during the second week of therapy on the circulating ANC (P = .01). The bone marrow neutrophil storage and proliferative pools were also significantly increased in newborn rats treated with rrSCF + rhG-CSF versus rhG-CSF (P = .02). The bone marrow and liver/spleen CFU-GM pool was unchanged; however, the CFU-GM proliferative rates were significantly increased in the rrSCF + rhG-CSF group (P = .04). rrSCF also induced a significant increase in the bone marrow and liver/spleen mast cell pool (P = .002). Lastly, rrSCF x 14 days +/- rhG-CSF significantly reduced the mortality rate at 48 and 120 hours after experimental group B streptococcus sepsis (P = .03 and .05, respectively). These data suggest that combination SCF + G-CSF therapy compared with G-CSF alone significantly increases the neonatal rat peripheral neutrophil count, bone marrow myeloid pools and proliferative rates, and induces a reduction in the mortality rate during experimental bacterial sepsis. SCF therapy may have future potential applications in the modulation of human neonatal hematopoiesis and host defense.

Animals↗

PubMed1988

Haematological abnormalities and reference intervals in the elderly. A cross-sectional comparative study of three urban Swedish population samples aged 70, 75 and 81 years.

We compared three representative population samples aged 70 (n = 312), 75 (n = 486) and 81 (n = 404) years. Anaemia defined either as blood haemoglobin concentration below the corresponding health-related lower reference limits or according to WHO was, in the total study groups, more common at age 81. Anaemia of unknown cause was rare, but somewhat more common at age 81. Mean blood haemoglobin concentrations in subsamples without definable disorders were significantly lower at age 81 than at ages 70 and 75, whereas the lower health-related reference limits did not differ significantly. White blood cell counts tended to be lower and iron deficiency was somewhat more common at age 81. No significant differences were found in platelet counts or in the prevalence of low plasma cobalamin concentrations. These results indicate a fall in blood haemoglobin with advancing age. The arbitrary WHO criteria for anaemia are clinically applicable at age 70-81.

Age Factors↗

PubMed1984

The effect of previous transection on quantitative estimates of the preganglionic neurones projecting in the cervical sympathetic trunk of the guinea-pig and the cat made by retrograde labelling of damaged axons by horseradish peroxidase.

The numbers of sympathetic preganglionic neurones in the upper thoracic spinal cord retrogradely labelled after application of horseradish peroxidase to the severed axons of the cervical sympathetic trunk have been determined in guinea-pigs and in cats. Neuronal counts were made on both sides of spinal segments C8 to T10 in control animals and in others in which the enzyme was applied 5-8 days after the left cervical sympathetic trunk had been transected. Labelling was not significantly different between sides of unoperated animals, but after previous cervical trunk section labelling was always less, by from 5 to 100% of that on the control side. The results were not significantly different if the application of horseradish peroxidase was made close to the original lesion or 1 cm more proximal along the nerve trunk. Analysis of the dimensions of labelled cells suggested that the deficit in labelling was greatest for small preganglionic neurones. These findings do not support an earlier report that retrograde labelling from regenerating axons is enhanced at this stage after axotomy, and the possible reasons for the discrepancy in results are discussed. It is suggested that labelling of cell bodies may not be restricted if intact axon sprouts are exposed to horseradish peroxidase. Reduced labelling from axons at the time of a second transection might, at least in part, be due to axon atrophy, emphasizing limitations to labelling when horseradish peroxidase is applied to severed axons of fine diameter. The high numbers and reproducibility of our control data enabled estimates to be made of the segmental distribution of the cell bodies or origin of the axons of the cervical sympathetic trunk in both species.

Animals↗

PubMed2001

Determination of somatic cells in milk by solid phase cytometry.

The somatic cell count of milk is routinely determined by the fluoro-opto-electron method and sometimes by the direct epifluorescent filter technique (DEFT). This paper investigates the potential of solid phase cytometry (SPC), a novel technique combining aspects of both the fluoro-opto-electronic method and epifluorescence microscopy for somatic cell counting. In SPC, cells are retained on a membrane filter, fluorescently labelled and automatically detected on the entire membrane filter by means of a laser scanning instrument ChemScan). Fluorescent spots can be visually inspected by an epifluorescence microscope with a computer-driven moving stage. The performance of SPC was compared with that of the fluoro-opto-electronic method using a Fossomatic 360 instrument for 68 milk samples with varying somatic cell counts (10(3)-10(6)/ml). The sample throughput and repeatability of SPC were inferior to those of' the Fossomatic method and statistical analysis of the method comparison data using the approach of J. M. Bland & D. G. Altman (The Lancet 1986 February 8 pp 307-310) revealed a poor comparability between the two methods. Moreover, problems of milk filterability and the interference of fluorescent particles presently hamper the routine application of SPC. Nevertheless, this method represents the first example of the application of SPC to milk.

Animals↗

PubMed1992

A calibrator for the determination of the erythrocyte concentration in electronic blood cell counting.

A stable calibrator, suitable for the quality control of electronic counting of erythrocytes, has been developed. The assigned concentration is determined by the cooperation of six laboratories using various types of blood cell counting instruments. The performance of the instruments was checked prior to operation. If required, corrections were applied, and deviations were taken into account. The suitability of the calibrator was tested in practice during a period of several years. The applicability of the preparation as a calibrator for use in routine practice was tested in 14 laboratories.

Calibration↗

PubMed1983

Changes of the activity of acid phosphatase and beta-glucuronidase in isolated platelets of whole-body irradiated rats.

In the work presented here we paid our attention to studying changes of the activity of two lysosomal enzymes-acid phosphatase and beta-glucuronidase in platelets after whole-body irradiation and application of certain clinically applied antifibrinolytics (Antilysin Spofa and epsilon-aminocaproic acid) on the activity of these enzymes in isolated platelets. From the results it is obvious that the activity of acid phosphatase and beta-glucuronidase increases in platelets of irradiated rats with a maximum on the 11th day after irradiation. In the same time interval after irradiation the most remarkable thrombocytopenia was observed. The application of Antilysin or EACA to intact, non-irradiated rats affects remarkably neither the activity of the followed enzymes nor the count of platelets. A statistically significant thrombocytopenia, in a comparison with controls was found in whole-body irradiated rats who received EACA or Antilysin. The postirradiation increase of the level of acid phosphatase in platelets exerts a similar character in rats who received EACA or Antilysin as in those who were only irradiated with a difference that the increase of the level of this enzyme was essentially lower in all the time intervals of interest. The beta-glucuronidase level in platelets of rats, who received EACA, was essentially unchanged as compared to controls. The application of Antilysin altered remarkably the level of beta-glucuronidase in platelets of whole-body irradiated rats in a comparison with a level of this enzyme in platelets of rats who received no Antilysin. Most remarkable changes were observed in latter time intervals (11th and 15th day) when the beta-glucuronidase level in platelets of rats after the application of Antilysin was remarkably higher than that in solely irradiated rats.

Acid Phosphatase↗

PubMed2003

Dip application of phosphates and marinade mix on shelf life of vacuum-packaged chicken breast fillets.

The effects of the dip application of a marinade on the shelf life of refrigerated raw vacuum-packaged broiler breast fillets simulating a retail product were evaluated over a 28-day period (at 0, 7, 14, 21, and 28 days) through enumeration of aerobic and anaerobic bacteria and total coliforms. Treatment effectively extended shelf life by ca. 14 days (with shelf lives of 28 and 14 days for dipped and control samples, respectively). Coliform growth was inhibited by the treatment, as indicated by counts of 1.6 and 7.4 log CFU/g for dipped and control fillets, respectively, on day 28. Initial coliform counts were similar for both types of fillets on day 0, averaging 2.0 log CFU/g. The data obtained in this study suggest that the dip application of a phosphate-spice marinade prior to the vacuum packaging of raw poultry may be an effective means of extending the shelf life and possibly improving the safety of raw poultry while providing value-added convenience and flavor.

Animals↗

PubMed1992

Examination of bacterial characteristics of anaerobic membrane bioreactors in three pilot-scale plants for treating low-strength wastewater by application of the colony-forming-curve analysis method.

Characteristic sludge ecosystems arising in anaerobic membrane bioreactors of three pilot-scale plants treating low-strength (less than 1 g of biological oxygen demand per liter) sewage or soybean-processing wastewater were examined by analysis of the colony-forming-curves (CFC) obtained by counting colonies at suitable intervals. The wastewaters, containing high amounts of suspended solids (SS) (SS/chemical oxygen demand ratio, 0.51 to 0.80), were treated by using two types of bioreactors: (i) a hydrolyzation reactor for solubilization and acidification of SS in wastewater and (ii) a methane fermentation reactor for producing methane. The colony counts for the two sewage treatment plants continued to increase even after 3 weeks of incubation, whereas those for soybean-processing wastewater reached an approximately constant level within 3 weeks of incubation. The CFCs were analyzed by correlating the rate of colony appearance on roll tubes with the physiological types of bacteria present in the bioreactors. It was found that there were large numbers of slow-colony-forming anaerobic bacteria within the bioreactors and that the viable populations consisted of a few groups with different growth rates. It is considered that the slow-growing colonies appearing after 10 days of incubation were the dominant microflora in the sewage treated by hydrolyzation reactors. In particular, highly concentrated sludge (30.0 g of mixed-liquor volatile SS per liter) retained by the membrane separation module contained a large number of such bacteria. Slow-growing colonies of these bacteria could be counted by using a sludge extract medium prepared from only the supernatant of autoclaved sludge. In addition, the highest colony counts were almost always obtained with the sludge extract medium, meaning that most of the anaerobic bacteria in these sludges have complex nutrient requirements for growth. This report also indicates the usefulness of application of the CFC analysis method to the study of bacterial populations of anaerobic treatment systems.

Journal Article↗

PubMed2004

Eubacterial 16S-rDNA amplicon profiling: a rapid technique for comparison and differentiation of heterotrophic plate count communities from drinking water.

Determination of the heterotrophic plate count (HPC) is commonly used as a surrogate to assess the general microbial water quality in drinking water. For routine monitoring applications, the HPC is investigated in a quantitative way. However, qualitative data about the HPC bacterial community composition and/or population dynamics are required for particular situations. In order to provide fast and efficient qualitative approaches, molecular biological DNA profiling techniques seem to be suitable tools for the analysis of the total HPC community composition. In this work a DNA profiling technique is presented, which was recently demonstrated by our group to have potential for the rapid qualitative comparison and differentiation of HPC communities from raw and drinking water. The presented approach consists of a polymerase chain reaction (PCR)-based denaturing gradient gel electrophoresis (DGGE) for the generation of 16S-rDNA amplicon fingerprints from whole HPC community DNA extracts. In the context of this proceeding, the methodical background is presented and possible scientific merits as well as potential water management applications are discussed. Selected examples of (i) the demonstration of selective growth of HPC populations on different media and the comparison to the total in situ drinking water eubacterial community, (ii) the screening for HPC community variations at different locations of a drinking water distribution system, and (iii) the influence assessment on groundwater HPC communities by an infiltrating treated sewage effluent (bacterial source tracking) are given.

Colony Count, Microbial↗