Male infertility is a relatively common condition affecting approximately 1 in 20 of the male population. DNA fragmentation is an important factor in the etiology of male infertility. Men with high DNA fragmentation levels have significantly lower odds of conceiving, naturally or through procedures such as intrauterine insemination and IVF. The most common contributing factor of male infertility is smoking. Studies have shown that smoking intensity is positively associated with job demands and stress. Therefore, we believe that work stress increases the nicotine-dependent thus causing lower male fertility rate. As proper protamine to histone ratio is essential to produce viable sperm, smoking is strongly suspected to reduce sperm viability through histone-to-protamine transition abnormalities. These abnormalities, results in sperm with high DNA damage when exposed to excessive free radical. This present study was undertaken to evaluate the relationship of work stress, smoking and sperm quality. A total of 210 infertile patients attending Medical Assisted Contraceptive Clinic (MAC), UKMMC were selected for the study. Smoking status and stress level of patients were collected after obtaining relevant consent. Histone-to-protamine ratio was acquired using Aniline Blue staining and Chromomycin A3 staining respectively. Sperm DNA fragmentation was estimated using Comet Assay. Result revealed that smokers tend to be more stressful (r = .446, p <. 001). The result showed a significantly increased level of histone (r = .385, p <. 001) and incomplete protamination (r = .492, p <. 001) in smokers. The imbalance of histone-to-protamine ratio lead to increase of DNA damage. All the data were analyzed using SPSS version 20.0. Result revealed that patients who smoke are more stressful at work. Higher proportion of abnormal sperm histone to protamine ratio were found among smokers suggesting that cigarette smoking may inversely affect male fertility.
male ; stress ; smoking ; sperm quality

OBJECTIVETo discuss how some pre-analysis processes influence the results of semen analysis and how to minimize their influence on the accuracy of laboratory results based on the concept of total quality management (TQM).

METHODSWe conducted semen quality analyses for 21 male volunteers, who had abstained from tobacco and alcohol for over 72 days for the purpose of fertilization, before and after the abstinence, and obtained their seminal parameters at 0.5, 1, 2 and 3 hours after semen sample collection.

RESULTSSperm concentration, sperm motility and the percentage of grade a + b sperm were significantly higher after the abstinence of tobacco and alcohol than before (P < 0.01). With the lengthening of post-ejaculation time, there was a significant decrease in sperm motility and the percentage of grade a + b sperm (P < 0.05), but not in sperm concentration (P > 0.05).

CONCLUSIONA lot of factors may affect the results of semen analysis, including the subjects' habits of drinking and smoking and the length of time after semen collection. Therefore, every procedure of semen analysis has to be dealt with very carefully so as to meet the requirements of TQM and achieve most reliable results for clinical use.

Adult ; Humans ; Male ; Quality Control ; Semen Analysis ; methods ; standards ; Smoking Cessation ; Sperm Count ; Sperm Motility ; Temperance

OBJECTIVETo evaluate the accuracy and precision of 4 methods including Hemacytometer, Makler chamber, Cell-VU chamber, and computer-aided semen analysis for determining sperm concentration.

METHODSLatex bead solutions with concentrations known as (35 +/- 5) x 10(6)/ml and (18.0 +/- 2.5) x 10(6)/ml and semen samples (n = 54) were counted by the above 4 methods and the results were then compared.

RESULTSMean bead concentrations for Hemacytometer, Makler, Cell-VU chambers and CASA were (44.84 +/- 4.86), (52.36 +/- 7.78), (39.70 +/- 4.76), (28.53 +/- 2.06) x 10(6)/ml respectively for the standard solution containing (35 +/- 5) x 10(6)/ml, and (21.04 +/- 1.87), (24.54 +/- 3.67), (19.09 +/- 2.02), (14.62 +/- 0.95) x 10(6)/ml respectively for a standard solution containing (18 +/- 2.5) x 10(6)/ml. The results of Cell-VU chamber were consistently similar and close to the standard solutions, while those of Hemacytometer, Makler chambers were overestimated, and those of CASA were underestimated. The coefficients of variation for Hemacytometer, Makler, Cell-VU chambers and CASA were 10.81%, 14.86%, 12.80%, and 7.22% respectively for a higher standard solution, while 8.89%, 14.96%, 10.58%, and 6.50% respectively for a lower standard solution. CASA has the lowest CV%. When semen samples were counted, the results of Hemacytometer, Makler, Cell-VU chambers and CASA were (76.98 +/- 59.90), (63.89 +/- 53.84), (45.28 +/- 34.52), (41.96 +/- 31.93) x 10(6)/ml respectively. There wasn't any significant difference either between Cell-VU chamber and CASA (P = 0.71), or between Hemacytometer and Makler chamber (P = 0.14), while there was significant difference between Cell-VU chamber or CASA and Hemacytometer or Makler chamber (P < 0.05 or P <0.01).

CONCLUSIONWhen counting semen sample, there wasnt any significant difference between Cell-VU chamber and CASA. Each laboratory can select its own proper method for manual or computer-aided analysis.

Diagnosis, Computer-Assisted ; Humans ; Male ; Oligospermia ; diagnosis ; Quality Control ; Sperm Count ; methods ; Sperm Motility

The semen analysis is one, if not the most, important and widely used clinical laboratory test to evaluate the fertility potential of the male. However, recent reports have suggested that the semen analysis is unreliable. Quality control in the andrology laboratory is often seen as problematic, and many laboratories do not routinely employ QC procedures in semen testing. Quality assurance is an often overlooked and unappreciated aspect of overall quality laboratory performance. External proficiency testing programs in andrology are not universally accepted, and the results from the few programs currently available demonstrate huge variations between laboratories. Numerous different standards and criteria are being used by andrology laboratories, making it difficult if not impossible to compare results from one laboratory to another. However, reliable semen analyses can be obtained by following several recommendations: (1) all laboratories performing the semen analysis should adopt universally accepted performance standards and criteria, (2) all laboratories performing this test should participate in external proficiency testing programs, (3) andrology laboratories should implement effective internal quality control and quality assurance programs to ensure that the results reported are accurate and reproducible, and (4) physicians should only refer their patients to, or accept semen analysis results from, laboratories that have stringently followed these recommendations.
Clinical Laboratory Techniques ; standards ; Humans ; Male ; Quality Control ; Reproducibility of Results ; Semen ; Sperm Count ; standards ; Sperm Motility

Semen analysis is characterized by high levels of intra- and inter-laboratory variability, due to a low level of standardization, high subjectivity of the assessments, and problems with automated procedures. To improve consistency of laboratory results, quality control and training of technicians are important requisites. The goals of this study are to evaluate the results of an external quality control (EQC) program and standardized training by ESHRE Basic Semen Analysis Courses (BSAC) on the variability in manual assessments of semen parameters. We performed retrospective analyses of (1) the interlaboratory variability in the Dutch EQC program and (2) the interobserver variability in BSACs for concentration, motility, and morphology assessments. EQC data showed that the interlaboratory coefficient of variation (CV) for concentration assessment decreased (range from 24.0%-97.5% to 12.7%-20.9%) but not for morphology and motility assessments. Concentration variability was lower if improved Neubauer hemocytometers were used. Morphology assessment showed highest CVs (up to 375.0%), with many outliers in the period of 2007-2014. During BSAC, a significant reduction of interobserver variability could be established for all parameters (P < 0.05). The absence of an effect in the EQC program for motility and morphology might be explained by respectively the facts that motility assessment was introduced relatively late in the EQC program (since 2013) and that criteria for morphology assessment changed in time. BSAC results might have been influenced by the pretraining level of participants and the influence of external factors. Both EQC and training show positive effects on reducing variability. Increased willingness by laboratories to change their methods toward standards may lead to further improvements.
Humans ; Netherlands ; Quality Control ; Retrospective Studies ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility

OBJECTIVETo establish a method for internal quality control (IQC) of sperm concentration test in the laboratory.

METHODSWe set the concentrations of frozen semen at 20 x 10(6) and 80 x 10(6) as low and high concentrations of putative IQC products, with QC-BEADSTM quality control beads (QCBs) as the control. Using the double-blind method, four technicians determined the sperm concentrations of the IQC products and QCBs by computer-assisted sperm analysis, and drew a quality control chart (Xbar chart and Sbar chart) for each product. Through a month of continuous detection, we calculated and compared the intra- and inter-batch coefficients of variation (CV%) of the quality control products of high and low concentrations.

RESULTSThe intra-batch coefficients of variation of the assumed IQC products of high and low concentrations were CV3.5% and CV2.4%, and their inter-batch coefficients of variation were CV10.2% and CV9.6%. The intra-batch coefficients of variation of the QCBs of high and low concentrations were CV5.1% and CV7.1%, and their inter-batch coefficients of variation were CV7.1% and CV8%. The intra-batch coefficients of variation of both IQC products and QCBs of high and low concentrations were <10%, and their inter-batch coefficients of variation were <15%, which conformed to Levey-Jennings quality control principles and achieved IQC purposes. No significant differences were found in either intra- or inter-batch coefficients of variation between the IQC products and QCBs of high and low concentrations (P>0.05), indicating that assumed IQC products can replace QCBs for internal quality control in the laboratory.

CONCLUSIONThe IQC method we established for determining sperm concentration is simple, feasible and reliable.

Double-Blind Method ; Humans ; Male ; Quality Control ; Semen Analysis ; methods ; standards ; Semen Preservation ; Sperm Count ; Sperm Motility ; Spermatozoa

OBJECTIVETo investigate and analyze the results of the determination of sperm concentration, fructose concentration, alpha-glucosidase and acid phosphatase (ACP) activities in the seminal plasma from different hospitals in the city of Nanjing, so as to provide a basis for the external quality control (EQC) of semen analysis within Jiangsu Province or even the whole country.

METHODSEight samples of quality control products for low and high concentrations sperm count, fructose, alpha-glucosidase and ACP determination were prepared and divided, each detected for the sperm concentration, fructose, alpha-glucosidase and ACP activity, and the coefficient variances (CVs) were calculated. The products were then distributed to 11 hospitals in the city, and the results were collected and analyzed. In addition, the total relative errors (REs) for each product was calculated based on the results after dividing as reference values.

RESULTSThe CVs from the 8 samples after dividing were 3.83% - 11.16%. Collected from the 11 hospitals attending EQC were 11 reports of the results of sperm concentration, and 5 the results of fructose, alpha-glucosidase and ACP in seminal plasma. Among the results from different laboratories, those of fructose determination showed the minimal difference (CVs: 8.99% and 3.95% for low and high concentrations, respectively) , next came alpha-glucosidase (CVs: 16.66% and 18.41% for low and high activities, respectively), and ACP determination showed the maximal difference (CVs: 54.12% and 65.58% for low and high activities, respectively). Moreover, the same trend was observed in RE values, as shown in the total REs, which were 11.99% (low concentration) and 20.31% (high concentration) for the determination of fructose in seminal plasma, 22.92% and 27.26% for alpha-glucosidase, 7.34% and 318.35% for ACP in different laboratories, and the maximal RE value was detected in the result of the high-activity ACP sample. Of the 11 hospitals, 6 determined sperm concentration with the computer-assisted semen analysis (CASA) system, and the other 5 with the modified hemocytometer. RE values (148.47% and 187.59% for low and high concentration samples, respectively) and sperm concentrations ([62.74 +/- 16.63] x 10(6)/ml and [163.32 +/- 36.24] x 10(6)/ml) counted with the hemocytometer were significantly higher than those with the CASA system (REs 13.97% and 10.48%; sperm concentrations [24.88 +/- 4.16] x 10(6)/ml and [54.24 +/-23.06] x 10(6)/ml ).

CONCLUSIONThe methods of seminal alpha-glucosidase and fructose determination were relatively stable in current andrology laboratories, and the variance range could be accepted. However, the method of seminal ACP determination might be unadaptable to clinical application, and needs to be further improved. Hemocytometer, which significantly overestimated sperm concentration, could not be applied to the assay of sperm concentration.

Acid Phosphatase ; analysis ; China ; Humans ; Male ; Quality Control ; Semen ; enzymology ; Sperm Count ; standards ; Sperm Motility ; alpha-Glucosidases ; analysis

Semen analysis is a basic test to evaluate male reproductive function. In recent years, urgent needs for the standardization of semen analysis have been emphasized among andrologists worldwide. This review discusses the standardization and quality control (QC) for the analysis of sperm quality parameters, including sperm concentration, motility and morphology. The key to sperm concentration analysis is the standardization of sperm-counting chamber, thus Cell-VU chamber may be the first choice. The analysis of sperm motility and morphology is too subjective to be reliable. Therefore, the computer-aided semen analysis (CASA) system may be the final selection. QC of semen analysis mainly lies in the selection of QC materials and the administration of external QC and internal QC. Meanwhile, the charts and arithmetic methods should be established to monitor QC of semen analysis.
Humans ; Male ; Quality Control ; Reference Standards ; Semen ; cytology ; Sperm Count ; instrumentation ; standards ; statistics & numerical data ; Sperm Motility ; Spermatozoa ; cytology ; physiology

No abstract available.
Animals ; Embryonic Structures* ; Fertilization in Vitro* ; Fetal Blood* ; Humans* ; Mice* ; Quality Control* ; Sperm Motility* ; Spermatozoa*

Clinical Laboratory Techniques ; standards ; Humans ; Male ; Quality Control ; Semen ; Sperm Count ; methods ; standards