Diagnosis of soil-transmitted helminths using the Kato-Katz technique: What is the influence of stirring, storage time and storage temperature on stool sample egg counts?

Background Soil-transmitted helminths infect about one fifth of the world’s population and have a negative impact on health. The Kato-Katz technique is the recommended method to detect soil-transmitted helminth eggs in stool samples, particularly in programmatic settings. However, some questions in its procedure remain. Our study aimed to investigate the effect of storage time, storage temperature and stirring of stool samples on fecal egg counts (FECs). Methodology/Principal findings In the framework of a clinical trial on Pemba Island, United Republic of Tanzania, 488 stool samples were collected from schoolchildren. These samples were evaluated in three experiments. In the first experiment (n = 92), two Kato-Katz slides were prepared from the same stool sample, one was stored at room temperature, the other in a refrigerator for 50 hours, and each slide was analyzed at nine time points (20, 50, 80, 110, 140 minutes, 18, 26, 42 and 50 hours). In the second experiment (n = 340), whole stool samples were split into two, one part was stored at room temperature, and the other part was put in a refrigerator for 48 hours. From each part one Kato-Katz slide was prepared and analyzed at three time points over two days (0, 24 and 48 hours). In the third experiment (n = 56), whole stool samples where stirred for 15 seconds six times and at each time point a Kato-Katz slide was prepared and analyzed. Mean hookworm FECs of Kato-Katz slides stored at room temperature steadily decreased following slide preparation. After two hours, mean hookworm FECs decreased from 22 to 16, whereas no reduction was observed if Kato-Katz slides were stored in the refrigerator (19 vs 21). The time x storage interaction effect was statistically significant (coefficient 0.26, 95% CI: 0.17 to 0.35, p < 0.0001). After 24 hours mean hookworm FECs dropped close to zero, irrespective of the storage condition. Whole stool samples stored at room temperature for one day resulted in a mean hookworm FEC decrease of 23% (p < 0.0001), compared to a 13% reduction (p < 0.0001) if samples were stored in the refrigerator. Fecal egg counts of A. lumbricoides and T. trichiura remained stable over time regardless of storage temperature of whole stool samples. Finally, we found a significant reduction of the variation of hookworm and T. trichiura eggs with increasing rounds of stirring the sample, but not for A. lumbricoides. For hookworm we observed a simultaneous decrease in mean FECs, making it difficult to draw recommendations on stirring samples. Conclusions/Significance Our findings suggest that stool samples (i) should be analyzed on the day of collection and (ii) should be analyzed between 20–30 minutes after slide preparation; if that is not possible, Kato-Katz slides can be stored in a refrigerator for a maximum of 110 minutes.

Download Full-text

Storage Time and Temperature on the Sensory Properties Broccoli

Foods ◽

10.3390/foods8050162 ◽

2019 ◽

Vol 8 (5) ◽

pp. 162 ◽

Cited By ~ 2

Author(s):

Robert Pellegrino ◽

Jennifer Wheeler ◽

Carl E. Sams ◽

Curtis R. Luckett

Keyword(s):

Storage Time ◽

Storage Temperature ◽

Consumer Acceptance ◽

Biochemical Processes ◽

Time Points ◽

Quality Degradation ◽

Two Temperatures ◽

Sensory Changes ◽

Acceptance Tests ◽

Holding Temperature

Typically, broccoli arrives at the store within 7–14 days of harvest and is kept refrigerated until purchased or considered waste. To date, information has been limited on how this time on the shelf or storage temperature affects the sensory attributes that contribute to broccoli purchase or repurchase. In this study, 100 consumers performed acceptance tests and a check-all-that-apply (CATA) section to characterize sensory changes in two cultivars of broccoli (‘Diplomat’ and ‘Emerald Crown’) stored at two temperatures (0 °C and 4 °C) over five time points: 0, 14, 21, 28, and 42 days. Due to quality degradation during storage, the overall liking of broccoli decreased regardless of holding temperature and variety. This was in accordance with a decrease in sweetness and an increase in bitterness intensity. However, there were differences between varieties in which Diplomat had more sensory changes at higher temperatures and only Emerald showed negative changes to its appearance in color. Lastly, the CATA data revealed the attributes responsible for modulating the consumer acceptance of broccoli such as tastes, colors and flavors (e.g., grassy, musty, dirt-like). This information can be used to better inform shelf-life determinations of broccoli. Additionally, these changes in taste, odor, texture, and color can inform those interested in investigating the biochemical processes related to broccoli storage.

Download Full-text

Influence of storage time and temperature on the activity of urease

Bulgarian Chemical Communications ◽

10.34049/bcc.51.2.4536 ◽

2019 ◽

Vol 51 (2) ◽

pp. 159-163

Author(s):

B. Alev ◽

S. Tunali ◽

R. Yanardag ◽

A. Yarat

Keyword(s):

Room Temperature ◽

Storage Time ◽

Urease Activity ◽

Storage Temperature ◽

Practical Importance ◽

Storage Conditions ◽

Relative Activity ◽

Significant Loss ◽

Long Term Storage ◽

Activity Measurements

Enzymes are made of protein, that is why they are sensitive molecules and are affected by storage conditions. A small change in enzyme activity during storage may cause a big error in analysis results. The aim of the study was to evaluate the effects of storage time and temperature on urease activity. Urease solutions were prepared at different activities (from 100 to 2000 U/mL) and stored at room temperature, in the refrigerator (4°C), and in the deep freezer (-18°C and -80°C). Activity measurements were made at regular intervals until 28 days by the modified Weatherburn method. The relative activities of 100-1000 U/mL urease solutions stored at room temperature, 4, -18 or -80°C were 75% and below after 4 days. Twenty-eight days later, for 2000 U/mL urease solutions, only at room temperature, the relative activity was reduced to 37%, while at 4, -18 or -80°C, the relative activities were above 80%. Since urease can be maintained at 4°C for 28 days without significant loss of activity, it has practical importance. Low-activity urease solutions (such as 100-1000 U/mL) should not be stored at -18 or -80°C for short or long term storage, they should be stored at 4°C only for one day. Keywords: Urease activity, storage time, storage temperature

Download Full-text

Formulation of Buprenorphine for Sublingual Use in Neonates

The Journal of Pediatric Pharmacology and Therapeutics ◽

10.5863/1551-6776-16.4.281 ◽

2011 ◽

Vol 16 (4) ◽

pp. 281-284 ◽

Cited By ~ 1

Author(s):

Ellena A. Anagnostis ◽

Rania E. Sadaka ◽

Linda A. Sailor ◽

David E. Moody ◽

Kevin C. Dysart ◽

...

Keyword(s):

Room Temperature ◽

Storage Time ◽

Storage Temperature ◽

Sublingual Administration ◽

Tandem Mass ◽

Abstinence Syndrome ◽

Length Of Treatment ◽

The Mean ◽

The Stability ◽

Length Of Hospitalization

OBJECTIVES The only medication used sublingually in the neonate is buprenorphine for the treatment of neonatal abstinence syndrome (NAS). Compared with morphine, buprenorphine reduces the length of treatment and length of hospitalization in neonates treated for NAS. The objective of this study was to characterize the stability of ethanolic buprenorphine for sublingual administration. METHODS Buprenorphine solution was prepared and stored in amber glass source bottles at either 68°F to 77°F (20°C-25°C) or 36°F to 46°F (2.2°C-7.8°C). Samples were collected from each of these batches on days 0, 3, 7, 14, and 30. Additional samples were withdrawn at baseline from each batch and placed in oral dispensing syringes for 3 and 7 days. Buprenorphine concentration was assessed by liquid chromatography–electrospray ionization–tandem mass spectrometry. RESULTS Neither storage temperature (p=0.65) nor storage time (p=0.24) significantly affected buprenorphine concentrations. All of the mean concentrations, regardless of storage temperature, were above 95% of the labeled concentration, and the potency was maintained for samples stored either in the original amber glass source bottles or in oral syringes. CONCLUSIONS An ethanolic buprenorphine solution is stable at room temperature for 30 days.

Download Full-text

Prevalence, Intensity of Soil-Transmitted Helminths, and Factors Associated with Infection: Importance in Control Program with Ivermectin and Albendazole in Eastern Côte d’Ivoire

Journal of Tropical Medicine ◽

10.1155/2019/7658594 ◽

2019 ◽

Vol 2019 ◽

pp. 1-10 ◽

Cited By ~ 2

Author(s):

Agodio Loukouri ◽

Aboulaye Méité ◽

Olivier K. Kouadio ◽

Norbert N. Djè ◽

Gotré Trayé-Bi ◽

...

Keyword(s):

Control Program ◽

The Other ◽

Intestinal Helminths ◽

Open Defecation ◽

Soil Transmitted Helminths ◽

Lack Of Information ◽

Chi Squared ◽

Successful Control ◽

Stool Samples

Evaluation of soil-transmitted helminths (STHs) and implementation of additional interventions are required in the region of a filariasis control program, given that antifilariasis drugs also have a beneficial effect on STHs. Thus, this study determines the extensive epidemiology of STHs to improve their successful control. Stool samples were analyzed using the Kato-Katz method. Chi-squared and Kruskal-Wallis tests were used to measure differences in infection rates and intensities, respectively, and logistic regression identified the risks of infection. The main intestinal helminths (A. lumbricoides, hookworm [N. americanus],S. mansoni,andT. trichiura) were found in the population. The overall prevalence of STHs was 19.5%. The prevalence of hookworm, the predominant species, ranged from 2% (n=6) to 28% (n=97). The overall prevalence of the other intestinal helminths was less than 6% (n=18). Intensity of hookworm was mostly light with a range from 1.6% (n=5) to 25.9% (n=90). However, the intensity of the species was significantly greater in Soribadougou compared to the other localities. Heavy infection was found in old children and adults but not in young children. Open defecation (OR=3.23, p≤0.05), dog/cat raising (OR=1.94, p≤0.05), farming (OR=14.10, p≤0.05), and irrigated culture (OR=3.23, p≤0.05) were positively associated with hookworm. It was observed that the participants missed the follow-up examinations due to trip (32.7%) or misunderstanding (15%) and lack of information (11.8%) of the purpose of the survey. Thus, to sustain the control of STHs, the MDA program should target the entire community and add education about the use of toilets, best practices of farming, and dog/cat raising.

Download Full-text

Factors affecting the accuracy and reliability of the measurement of anti-Müllerian hormone concentration in the clinic

Journal of International Medical Research ◽

10.1177/03000605211016161 ◽

2021 ◽

Vol 49 (5) ◽

pp. 030006052110161

Author(s):

Yun-Xing Fu ◽

Hui Wang ◽

Ting Hu ◽

Fei-Miao Wang ◽

Rong Hu

Keyword(s):

Gnrh Agonist ◽

Room Temperature ◽

Storage Time ◽

Storage Temperature ◽

Oral Contraceptive Pill ◽

Microtitre Plate ◽

Serum Samples ◽

Factors Affecting ◽

Measurement Placement ◽

Gonadotrophin Releasing Hormone

Objective We aimed to identify the factors that influence serum anti-Müllerian hormone (AMH) concentration measurements. Methods We collected serum samples between May and September 2018 and compared the effect on AMH concentration measured by ELISA of conditions including venepuncture, storage time, storage temperature, locations of the reaction microplate, and the use of the oral contraceptive pill and gonadotrophin-releasing hormone (GnRH). Results AMH concentration was not affected by food intake but was affected by haemolysis. It was also much higher in samples on the edge of the ELISA microtitre plate. AMH concentration increased after incubation at room temperature for 1 day, 4°C for 3 days, −20°C for 1 month and −40°C for 4 months, but no change occurred during storage at −80°C for 9 months. AMH concentration was high in patients following GnRH agonist treatment but was not affected by oral contraceptives. Conclusions No fasting is required prior to AMH measurement. Placement of serum samples on the edge of microtitre plates affects the results of the AMH ELISA. If serum samples cannot be assayed immediately, it is best to store them at −80°C. Basal AMH concentration cannot be used as a measure of ovarian reserve after GnRH agonist treatment.

Download Full-text

Effect of Storage Time and Temperature on Fresh Unprocessed Cord Blood.

Blood ◽

10.1182/blood.v106.11.5272.5272 ◽

2005 ◽

Vol 106 (11) ◽

pp. 5272-5272

Author(s):

Marcus R. Vowels ◽

Jessica Stylianou ◽

Leigh Mison

Keyword(s):

Cord Blood ◽

Room Temperature ◽

Storage Time ◽

Storage Temperature ◽

Haematopoietic Stem Cell ◽

Optimal Conditions ◽

Cd34 Cells ◽

Different Temperatures ◽

Thawing Process ◽

Temperature Ranges

Abstract Studies on the optimal conditions to store fresh cord blood (CB) in order to optimise haematopoietic stem cell (HSC) recovery prior to processing and cryopreservation have produced conflicting data. In this study we investigate the effects of time and temperature on CB HSC after collection. We also investigate whether the process of cryopreservation and thawing aggravates stress created by storage prior to cryopreservation. 30 CB units were collected and transported to the laboratory within 5 hours of collection. An aliquot of each CB unit was tested for nucleated cell count (NCC), CD34 count, CD34 viability and CFU-GEMM, and then frozen with 10% DMSO. Each CB unit was then separated into three 10 ml portions and incubated at 3 different temperatures (4–8°C, 21–24°C and 29–31°C). After 24, 36 or 48 hours, samples were taken and tested. An aliquot of each of the portions was then cryopreserved. After a minimum of 1 week, the cryopreserved samples were thawed and tested. For fresh samples incubated for 24 hrs or 36 hrs there was no difference over time when CB was incubated at 4–8°C, whereas, at temperatures above 20°C, there was either a trend or significant decrease in CD34 and CFU colonies with time. For thawed samples, viable CD34 and CFU colonies were decreased below baseline at either one or both 24 hr and 36 hr time points. This was particularly apparent for CFU colony measurements for samples kept at 29–31°C. At 48 hrs, at each of the temperature ranges (4–8°C, 21–24°C and 29–31°C), CFU were significantly lower for fresh (21.1, 16.8 and 14.0; baseline 31.0) and thawed (9.3, 8.5 and 3.0; baseline 2.7) samples and viable CD34 were significantly lower for thawed (2.0, 2.46 and 1.8; baseline 2.7) samples respectively (p<0.001). In conclusion, the best survival /recovery was seen with CB stored at 4–8° C and for ≥ 36 hrs. Temperatures above room temperature (21–24°C) appeared detrimental. The data also indicate that this damage becomes more evident when tested after the CB has gone through the cryopreservation and thawing process, and is even more evident the higher the temperature and the greater the time of storage prior to cryopreservation. These results have implications for quality and safety of CB stored for clinical use. Viable CD34 and CFU related to incubation time and temperature, tested pre- and post-cryopreservation. Viable CD34 cells x 10e6 CFU-GEMM /12,500 cells plated Storage temperature 4–8C° 21–24°C 29–31°C 4–8°C 21–24°C 29–31°C Baseline 3.4 3.4 3.4 20.9 20.9 20.9 Fresh 24 hrs 3.3 3.67 2.45 20.4 18.8 21.6 36 hrs 3.7 2.45 2.74 20.5 19.6 17.4 Frozen / Baseline 3.0 3.0 3.0 11.0 11.0 11.0 Thawed 24 hrs 2.7 2.8 2.5 11.8 11.8 6.1 36 hrs 2.5 2.84 2.1 10.6 10.1 4.7

Download Full-text

Improving IBD diagnosis and monitoring by understanding preanalytical, analytical and biological fecal calprotectin variability

Clinical Chemistry and Laboratory Medicine (CCLM) ◽

10.1515/cclm-2018-0134 ◽

2018 ◽

Vol 56 (11) ◽

pp. 1926-1935 ◽

Cited By ~ 15

Author(s):

Andrea Padoan ◽

Renata D’Incà ◽

Maria Luisa Scapellato ◽

Rudi De Bastiani ◽

Roberta Caccaro ◽

...

Keyword(s):

Room Temperature ◽

Storage Time ◽

Storage Temperature ◽

Clinical Effectiveness ◽

Fecal Calprotectin ◽

Time Interval ◽

Biological Variability ◽

Reference Change Value ◽

The Difference ◽

Clinically Significant

Abstract Background: The appropriate clinical use of fecal calprotectin (fCal) might be compromised by incomplete harmonization between assays and within- and between-subjects variability. Our aim was to investigate the analytical and biological variability of fCal in order to provide tools for interpreting fCal in the clinical setting. Methods: Experiments were conducted to investigate the effects of temperature and storage time on fCal. Thirty-nine controls were enrolled to verify biological variability, and a case-control study was conducted on 134 controls and 110 IBD patients to compare the clinical effectiveness of three different fCal assays: ELISA, CLIA and turbidimetry. Results: A 12% decline in fCal levels was observed within 24 h following stool collection irrespective of storage temperature. Samples were unstable following a longer storage time interval at room temperature. Within- and between-subjects fCal biological variability, at 31% and 72% respectively, resulted in a reference change value (RCV) in the region of 100%. fCal sensitivity in distinguishing between controls and IBD patients is satisfactory (68%), and the specificity high (93%) among young (<65 years), but not among older (≥65 years) subjects (ROC area: 0.584; 95% CI: 0.399–0.769). Among the young, assays have different optimal thresholds (120 μg/g for ELISA, 50 μg/g for CLIA and 100 μg/g for turbidimetry). Conclusions: We recommend a standardized preanalytical protocol for fCal, avoiding storage at room temperature for more than 24 h. Different cutoffs are recommended for different fCal assays. In monitoring, the difference between two consecutive measurements appears clinically significant when higher than 100%, the fCal biological variability-derived RCV.

Download Full-text

IDENTIFIKASI TELUR CACING NEMATODA USUS SOIL TRANSMITTED HELMINTH (STH) PADA MASYARAKAT DI PULAU LAE-LAE KOTA MAKASSAR

Lontara ◽

10.53861/lontarariset.v2i1.188 ◽

2021 ◽

Vol 2 (1) ◽

pp. 42-48

Author(s):

Anita Anita Anita ◽

Tuty Widyanti ◽

Effendy Rasiyanto ◽

Budiawan S.HI. Karim

Keyword(s):

Stool Sample ◽

Sampling Technique ◽

The Other ◽

Laboratory Observation ◽

Intestinal Nematode ◽

Positive Stool ◽

Faecal Samples ◽

Stool Samples ◽

Positive Stool Sample ◽

Nematode Worm

ABSTRACT Communities on Lae-Lae Island, Makassar City have sanitation facilities that are still poor and very limited with quality far from health standards. This causes people who live in these islands to face various health problems, one of which is the risk of being infected with eggs of intestinal nematode worms Soil-Transmitted Helminths (STH). This study aims to identify the eggs of the intestinal nematode worm Soil Transmitted Helminthes in the feces of people on Lae-Lae Island, Makassar City. This type of research is a laboratory observation with a purposive sampling technique of 10 stool samples. Based on the results of research that has been carried out on 10 faecal samples, it was found 1 positive stool sample for Trichuris trichura worm eggs with distinctive egg-shaped characteristics such as the shape of crock worm eggs or wine barrels and at both ends there are two mucoid plugs. The egg wall is brown from the color of the bile at both ends, it is clear, while the other 9 stool samples are negative the type of worm Trichuris trichura and 9 other samples were negative.

Download Full-text

The Changes Of Characteristic Fermented Milk During Room Temperature Storage

Jurnal Ilmu dan Teknologi Pangan (ITEPA) ◽

10.24843/itepa.2021.v10.i01.p11 ◽

2021 ◽

Vol 10 (1) ◽

pp. 119

Author(s):

Hidayanti Sukmaningrum ◽

Luh Putu Trisna Darmayanti ◽

Gusti Ayu Kadek Diah Puspawati

Keyword(s):

Sensory Evaluation ◽

Room Temperature ◽

Storage Time ◽

Storage Temperature ◽

Fermented Milk ◽

Milk Product ◽

Total Acid ◽

Room Temperature Storage ◽

Acid Ph ◽

Temperature Storage

Fermented milk is a functional food has beneficial to human health. The recommended concentration of probiotic bacteria to provide health benefits is 106-108 CFU/ml of product. The storage temperature is one of the factor that affected the characteristics of fermented milk. This research was conducted to determine the effect of storage time at room temperature to the characteristics of fermented milk product and determine the length of maximum storage at room temperature. The research design was Completely Randomized Design with storage time treatment at room temperature for 0, 2, 4, 6, 8, 10, 12, and 14 days. Each treatment was repeated 2 times so that 16 experimental units were obtainned. The variables that were observed included total LAB, total acid, pH, and sensory evaluation (aroma, taste, and overall acceptance). Data were analyzed with analysis of variance, if the treatment had an effect on the variables then followed by Duncan multiple range Test. The results showed that the treatment of fermented milk in room temperature storage significantly affected the total acid, pH, total LAB, and sensory evaluation (aroma, taste, and overall acceptance). The maximum storage of the room temperature was days with total acid of 0.5%, pH 3.63, total LAB 7.71 log cfu/ml or 5.12 x 107 cfu/ml, the sensory of aroma, taste, and overall acceptance was liked.

Download Full-text

Monostearin effects on the formation of precipitate in palm oil biodiesel and petroleum diesel blends with various storage temperature

E3S Web of Conferences ◽

10.1051/e3sconf/20185200026 ◽

2018 ◽

Vol 52 ◽

pp. 00026 ◽

Cited By ~ 1

Author(s):

Mohamad Aufar Ghaizani ◽

Imam Abdurrosyid ◽

Imam Paryanto ◽

Misri Gozan

Keyword(s):

Low Temperature ◽

Palm Oil ◽

Room Temperature ◽

Storage Time ◽

Storage Temperature ◽

Biodiesel Production ◽

Blend Ratio ◽

Early Results ◽

Biodiesel Blend ◽

Palm Oil Biodiesel

Biodiesel is one of the renewable energy forms that is on demand in Indonesia. In the biodiesel production process, impurities such as Saturated Monoglycerides (SMG) in the form of monostearin that precipitate at low temperature are commonly formed. This is caused by high Final Melting Temperature (FMT) of SMG. Formation of solid deposits when it reaches a temperature above Cloud Point (CP) is then unavoidable under these conditions. The use of palm oil biodiesel and petroleum diesel blends (BXX) with biodiesel blend ratio of 10% (B10), 20% (B20) and 30% (B30) accelerates precipation process which renders clogging on fuel filters. These works examined the effect monostearin content and temperature on the precipitation rate. Investigation is carried out at 15°C, 20°C, 25°C, and room temperature (30-33°C) with varying content of monostearin (0.4%, 0.7%, and 0.9%). Early results show that for 0.4% monostearin content in 100 ml B20, the amount of precipitate formed at temperature 15 oC was 31.3 mg. This value was higher than that at room temperature (6.3 mg) after 2 weeks storage time. This value was higher compared to B20 (21.5 mg). This indicates that as the biodiesel ratio in BXX become higher, the amount of FAME will increase which effect monostearin solubility in BXX at low temperature.

Download Full-text