Round Cells In Semen Research Articles

Mild coronavirus disease 2019: Results of a multicentric prospective study on severe acute respiratory syndrome coronavirus 2 presence in semen fractions and cells and its effects on semen parameters.

Severe acute respiratory syndrome coronavirus 2, the virus responsible for coronavirus disease 2019, affects multiple organs. The virus enters cells through angiotensin-converting enzyme-2 and host factors present in genital organs, leading to concern over virus shedding in semen and reproductive function. To investigate severe acute respiratory syndrome coronavirus 2 in semen from patients with a mild infection, identify the seminal infected cells, and explore the effect of the infection on sex hormones and semen parameters. Prospective study of 54 men with mild severe acute respiratory syndrome coronavirus 2 infection. Semen was collected at 7, 15, 30, 60, 90, 180, and 365 days after symptom onset, and severe acute respiratory syndrome coronavirus 2 RNA was measured in serum, saliva, urine, and semen. The presence of infectious severe acute respiratory syndrome coronavirus 2 in semen was assessed using Vero cell culture. Infected semen cells were identified using immunofluorescence against severe acute respiratory syndrome coronavirus 2 nucleoprotein antigen and cell markers. Semen characteristics as well as testosterone, inhibin B, luteinizing hormone, and follicle-stimulating hormone levels were determined. 11% of patients had at least one severe acute respiratory syndrome coronavirus 2 RNA-positive semen. One patient had viral semen shedding up to day 90 after infection onset, with replication-competent virus isolated from semen and 40% cell fraction at day 7. After sperm preparation, 90% fraction was severe acute respiratory syndrome coronavirus 2 RNA-positive at days 7 and 15. The swim-up fraction was positive only on day 7. In semen, nucleoprotein antigen was detected mainly in exfoliated epithelial cells and less frequently in Sertoli cells. Sperm count and motile sperm count were lower at day 30 than at day 7. Round cells in semen were increased during the acute phase. At days 7 and 15, sperm count and motile sperm count were lower in severe acute respiratory syndrome coronavirus 2 RNA-positive semen compared with negative semen, while semen volume and follicle-stimulating hormone levels were increased. Long-term follow-up shows no evidence of a detrimental effect on hormonal or semen characteristics. 11% of patients with mild coronavirus disease 2019 who were not hospitalized had severe acute respiratory syndrome coronavirus 2 excretions in semen, which persisted for up to 90 days in one patient. No germ cells appeared infected by the virus, but the detection of nucleoprotein antigen-positive epithelial semen cells and Sertoli cells suggests genital tract infection. Albeit infrequent, semen may contain the replication-competent virus during the acute phase with potential risk of severe acute respiratory syndrome coronavirus 2 transmissions during sexual contact and assisted reproduction procedures. The effect of mild coronavirus disease 2019 on spermatogenesis and reproductive hormones was moderate and reversible.

Diff-Quik: A reliable stain for rapid onsite evaluation and differentiation of round cells in semen

Objectives: Identification of immature germ cells and leukocytes and their differentiation pose difficulty in interpretation. In this study, Diff-Quik (DQ) and Papanicolaou (Pap) are used to differentiate between these two, as they are recommended by the WHO, cost-effective and are easy to use. Materials and Methods: This study attempts to differentiate between immature germ cells and leukocytes in semen; compare the usefulness of two stains –Pap and DQ; evaluate the possible associations between the total sperm count and the round cell count; the total sperm count and the presence of round cells and inflammatory cells; the occupation and the sperm concentration; and the occupation and the round cell count. Results: A total of 60 semen samples were included in the study. Two smears were made for each patient; DQ stain was used to stain the air-dried smears and Pap stain for alcohol-fixed smears. The whole procedure of DQ staining takes only 30 s. The smears were compared. There was good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells (P = 0.000). There was no association between total sperm count and round cell count (r = −0.086 and P = 0.520); and between the occupation and the sperm concentration or the round cell count. Conclusion: There was an inverse relationship between total sperm count and presence of immature germ cells (r = −0.291). There was no association between round cell count and presence of immature germ cells (r = 0.148 and P = 0.269), total sperm count and presence of inflammatory cells, round cell count, and presence of inflammatory cells. DQ stain is easy to use, convenient, and rapid. There is good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells. Total sperm count is independent of the round cell count or the presence of inflammatory cells/immature germ cells. In most of the cases, the round cells were the inflammatory cells. Morphological interpretation can be done even in non-liquefied samples.

O-136 Myeloperoxidase Kit as valuable alternative for LeucoScreen to differentiate round cells in semen samples

Abstract Study question Is Myeloperoxidase Kit, validated for blood but not for semen, a valuable alternative for Leucoscreen to differentiate round cells in semen? Summary answer Myeloperoxidase Kit is a non-toxic alternative for LeucoScreen to differentiate round cells in semen, with the advantage of assessing a 1:2 dilution of semen. What is known already As stated in the WHO laboratory manual, the routine assay of peroxidase activity is useful as screening technique for quantifying the leucocyte population in semen samples. The clinically relevant neutrophil leucocytes stain peroxidase positive and can hereby be differentiated from spermatids and spermatocytes. The commercially available kit, Leucoscreen (Fertipro, Beernem, Belgium) uses carcinogenic ortho-toluidine and was therefore withdrawn from the market and an alternative, LeucoScreen Plus, was suggested. A disadvantage of LeucoScreen Plus is the necessity to dilute the sample 1:6. Testing an alternative inexpensive, quick and not carcinogenic method to differentiate round cells without excessive dilutions, is worth investigating. Study design, size, duration Intra- and inter-variability of Myeloperoxidase Kit (Ral Diagnostics, MLS NV, Menen, Belgium) was assessed by evaluating 2 semen samples 10 times by one technician, and 10 semen samples by 2 technicians, respectively. The stability of the working solution for 5 days was tested on 10 semen samples. The accuracy was tested on 80 samples by comparing Myeloperoxidase Kit with LeucoScreen. The study was performed from July till November 2020. Participants/materials, setting, methods Working solution was made from the Myeloperoxidase Kit (3/5 bottles): 1 ml alphanaphtol solution, 4 ml pyronine solution and 1 droplet hydrogen peroxide. Semen samples brought to the Andrology Lab for diagnostic evaluation and presenting with round cells were included. To differentiate peroxidase positive leucocytes (dark pink with dark granules) from other round cells (unstained or pink), 10 µl solution was mixed with 10 µl of semen and assessed with phase contrast at 400x. Main results and the role of chance The intra-variability of round cell differentiation was acceptable with %CV 6.6% for sample 1 (mean ± SD: 30.8% ± 2.04%) and 4.1% for sample 2 (mean ± SD: 56.7% ± 2.31%), both lower than the 10% criterium. The inter-variability was acceptable according to Passing&Bablok (95%CI Intercept A: -1.25 to 0.36; 95%CI Slope B: 0.92 to 1.11) and Spearman rank correlation coefficient (r = 0.983 with P < 0.001). The stability of the working solution after 5 days, kept at room temperature during the day and at 4 °C at night, compared with freshly made working solution was adequate according to Passing&Bablok (95%CI Intercept A: -2.33 to 0.47; 95%CI Slope B: 0.85 to 1.17) and Spearman rank correlation coefficient (r = 0.991 with P < 0.0001). Also the accuracy could be accepted as the Bland-Altman plot shows a substantial agreement between Leucoscreen and Myeloperoxidase Kit (difference 0.6%) without a proportional bias. Limitations, reasons for caution Granules in neutrophils stain differently with Myeloperoxidase Kit, with Leucoscreen and with Leucoscreen Plus, which requires a short adaptation when starting with a new kit. Wider implications of the findings The use of Myeloperoxidase Kit to differentiate round cells in semen is cheap, non-toxic and easy. It is a valuable asset in the andrology lab because semen samples with low round cell concentrations can be counted in a 1:2 dilution and the working solution can be kept for 5 days. Trial registration number None

Significance of positive semen culture in relation to male infertility and the assisted reproductive technology process.

There are currently no WHO guidelines on the indications for semen culture; however, semen cultures are performed in the evaluation of male infertility and the assisted reproductive technology (ART) process. The relevance and significance of positive semen cultures is widely debated in the literature, with no current consensus on the usefulness of this test in relation to male infertility. We review the pathogenic mechanisms of potentially pathogenic bacteria, general bacteria, urethral flora, and skin flora on sperm parameters. We also present, possible routes of semen contamination, measures to reduce contamination, and the clinical significance of culture contamination. First, it is critical to distinguish round cells in semen as leukocytes from immature germ cells. Second, it is critical to distinguish leukocytospermia from infected semen in order to guide management.

Sperm DNA fragmentation as a result of ultra-endurance exercise training in male athletes.

Intensive sports practice seems to exert negative effects on semen parameters; in order to assess these effects, the objective of this study was to assess semen, including DNA fragmentation, and hormone parameters in elite triathletes. Twelve high-level triathletes preparing for a National Triathlon Championship participated in the study. The qualitative sperm parameters analysed were volume, sperm count, motility, morphology and DNA fragmentation; when needed, additional testing was performed. Assessed hormones were testosterone (T), cortisol (C) and testosterone-cortisol ratio (T/C). Maximum oxygen consumption and training characteristics were also assessed. Hormonal values and physical semen parameters were within normal ranges. DNA fragmentation showed high values (20.4±6.1%). Round cells in semen were higher than normal (2.8±1.5 million/ml), with the presence of macrophages. Correlations were found for several parameters: concentration of round cells positively correlated with progressive sperm motility (p=.01) and sperm morphology (p=.02); contrarily, the correlation found with DNA fragmentation was negative (p=.04). Sperm DNA fragmentation and the T/C ratio, however, were correlated in a positive manner (p=.03). As evidenced by the observed results, sperm DNA fragmentation is affected by high-level sports practice; therefore, high loads of endurance training could potentially interfere with the athlete's fertility potential.

Findings on sperm alterations and DNA fragmentation, nutritional, hormonal and antioxidant status in an elite triathlete. Case report

ObjectiveThe present case study analyzes semen quality, nutritional patterns, and hormonal and oxidative status of an international high-level triathlete with a low-volume, high-intensity training load. MethodThe athlete was 26 years old, having participated in competitions since he was 13 years old, and practiced professional triathlon for the last five years. The qualitative sperm parameters analyzed were volume, sperm count, motility, morphology, and DNA fragmentation (additional testing performed as needed). Salivary hormones measured were T, C, and T/C. Seminal plasma total antioxidant capacity was measured. Maximum oxygen consumption and training characteristics were assessed. To determine habitual food intake and its possible repercussion on oxidative status, a quali-quantitative frequency questionnaire of 136 different foods was used and subsequently analyzed using specific software. ResultsHormonal and physical semen parameters were within normal ranges. Sperm morphology and DNA fragmentation showed abnormal values (3.9% and 38.33%). Round cells in semen were higher than normal (2.3million/ml), with presence of macrophages. Apoptotic and necrotic events were observed. Total antioxidant capacity, although not compromised, was low. Dietetic intake was characterized by excess protein and appropriate overall antioxidant intake (with slight deficit and excess of some specific nutrients) according to recommended values. ConclusionsIn this high-intensity endurance athlete, sperm parameters, mainly sperm morphology and DNA fragmentation, are altered. Further knowledge is needed with regards nutritional antioxidant intake and other dietetic strategies oriented toward avoiding oxidative damage in semen of high-performance triathletes. Moreover, adequate nutritional strategies must be found and nutritional advice given to athletes so as to palliate or dampen the effects of exercise on semen quality.

Immature germ cells in semen - correlation with total sperm count and sperm motility

Background:Current data regarding infertility suggests that male factor contributes up to 30% of the total cases of infertility. Semen analysis reveals the presence of spermatozoa as well as a number of non-sperm cells, presently being mentioned in routine semen report as “round cells” without further differentiating them into leucocytes or immature germ cells.Aim:The aim of this work was to study a simple, cost-effective, and convenient method for differentiating the round cells in semen into immature germ cells and leucocytes and correlating them with total sperm counts and motility.Materials and Methods:Semen samples from 120 males, who had come for investigation for infertility, were collected, semen parameters recorded, and stained smears studied for different round cells. Statistical analysis of the data was done to correlate total sperm counts and sperm motility with the occurrence of immature germ cells and leucocytes. The average shedding of immature germ cells in different groups with normal and low sperm counts was compared. The clinical significance of “round cells” in semen and their differentiation into leucocytes and immature germ cells are discussed.Conclusions:Round cells in semen can be differentiated into immature germ cells and leucocytes using simple staining methods. The differential counts mentioned in a semen report give valuable and clinically relevant information. In this study, we observed a negative correlation between total count and immature germ cells, as well as sperm motility and shedding of immature germ cells. The latter was statistically significant with a P value 0.000.

Etiology of azoospermia in 100 consecutive nonvasectomized men

History was taken systematically for 100 azoospermic, nonvasectomized men referred consecutively to a Danish fertility clinic. The men were examined by ultrasound, and their blood samples were analyzed for karyotype, Y microdeletions, and cystic fibrosis transmembrane conductance regulator gene mutations. In 29% of patients, the condition could be explained by genetic abnormalities; in 22%, by diseases or external influence; and in 27%, by former cryptorchidism. The azoospermic condition remained unexplained in only 22%.

Flow cytometric analysis of semen preparation, and assessment of acrosome reaction, reactive oxygen species production and leucocyte contamination in subfertile men

Reactive oxygen species (ROS) production, poor acrosome reaction ability, and leucocyte and round cell contamination of semen are associated with diminished male fertility. In addition to the assessment of these variables, the number of abnormal and round cells in semen preparation was studied by flow cytometry. Men with varying degree of fertility despite normal concentrations and motility of spermatozoa were investigated. Group 1 consisted of semen donor candidates (n = 25), group 2 of patients with acceptable fertilization in vitro, but diminished fertility in vivo (n = 56), and group 3 of patients with poor (< 25%) fertilization in vitro (n = 32). The subfertile men in groups 2 and 3 had lower concentrations and poorer rapid progressive motility of spermatozoa than the men in group 1. Using flow cytometry, the number of events in the round cell gate in the Percoll-prepared semen sample was higher in the subfertile men than in the donors, independently of the differences in other semen parameters. However, there were no significant differences between the groups in the number of round cells in native semen by microscopic analysis, or in ROS production or acrosome reaction. In conclusion, flow cytometric assessment of round cells may be a useful method in the investigation of male fertility.

Immature germ cells in semen and their correlations with other semen parameters.

The significance of the presence of leukocytes and immature germ cells in semen and other parameters of semen is currently a subject of controversy. Semen from 572 subfertile patients was analyzed according to WHO criteria and leukocytes as well as immature germ cells were assessed by identifying the round cells in semen by peroxidase staining. Microbiological investigation was carried out in cases with leukocyte counts of >1,000,000/ml. It was found that as the concentration of spermatozoa decreased the rate of immature germ cells increased and this increase was accompanied by a decrease in motility and in the number of spermatozoa with normal morphology. As the sperm count increased, motility, number of spermatozoa with normal morphology and of immature germ cells also increased whereas an increase in sperm motility was accompanied by an increase in the percentage of spermatozoa with normal morphology. Microbiological investigations were negative in patients with semen leukocyte counts of >l,000,000/ml. Although it is possible to establish that the leukocyte and immature germ cell counts correlate with other parameters of semen, these correlations are not statistically significant. The most significant finding is that as the number of sperms decreases, the ratio of immature germ cells to total germ cells increases. While assessing immature germ cells instead of giving special attention to the number of immature germ cells in semen, the ratio of immature germ cells to total germ cells should be considered. The increase of leukocyte count in the semen of oligospermic patients may not always mean leukospermia.

Sperm morphology assessment using strict criteria and male fertility under in-vivo conditions of conception

The clinical significance of sperm morphology assessment according to very strict criteria was determined using semen samples of randomly chosen males from couples not submitted to assist procreation techniques, with a median duration of infertility of 4 years (range 1-17; n = 89). The relationships of sperm morphological properties to the results of standard sperm analysis, including the differentiation of round cells in semen by monoclonal antibodies and semen cultures, the testing of sperm functional capacity in vitro with the standardized sperm-cervical mucus penetration test (SCMPT) and the subsequent pregnancy rate under in-vivo conditions of conception, were evaluated in a prospective study. The quick staining method (DiffQuick(R) stain) for sperm morphology proved to be practical and suitable for routine use. The percentage of normal forms according to strict criteria ranged from 1 to 36%, with a median of 12%. Morphological findings were not markedly related to the medical history, but significant relationships between standard parameters of sperm analysis, in particular the sperm count, the progressive motility and standard sperm morphology, were found. The percentage of normal forms was not significantly associated with the microbial colonization of semen samples but was negatively related to high leukocyte rates. Semen samples with a higher percentage of normal spermatozoa (shown, for example, for >4,> 7 or >=14% normal) were significantly more frequent in cases of an adequate SCMPT. The subsequent pregnancy rate within an observation period of 12 months was 20.2%. The pregnancy rate under in-vivo conditions was significantly higher when semen samples had a better sperm morphology, with significant differences for thresholds at 4, 7 and 14% of strictly normal forms. Although sperm morphology is only one among a multiplicity of factors determining fertility, the results suggest that the evaluation of sperm morphology using strict criteria provides valuable information during basic infertility investigations.

Differentiation of round cells in semen by means of monoclonal antibodies and relationship with male fertility

International Journal of Gynecology & ObstetricsVolume 42, Issue 1 p. 86-87 Citations from the literature fertility, sterility Differentiation of round cells in semen by means of monoclonal antibodies and relationship with male fertility W. Eggert-Kruse, W. Eggert-KruseSearch for more papers by this authorA. Bellmann, A. BellmannSearch for more papers by this authorG. Rohr, G. RohrSearch for more papers by this authorW. Tilgen, W. TilgenSearch for more papers by this authorB. Runnebaum, B. RunnebaumSearch for more papers by this author W. Eggert-Kruse, W. Eggert-KruseSearch for more papers by this authorA. Bellmann, A. BellmannSearch for more papers by this authorG. Rohr, G. RohrSearch for more papers by this authorW. Tilgen, W. TilgenSearch for more papers by this authorB. Runnebaum, B. RunnebaumSearch for more papers by this author First published: July 1993 https://doi.org/10.1016/0020-7292(93)90472-9AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume42, Issue1July 1993Pages 86-87 RelatedInformation

The Correlation between Round Cells and White Blood Cells in the Semen

It is generally recognized that white blood cells in semen are detrimental to sperm function and fertility. Traditionally, pyospermia has been suspected if greater than 10 round cells per high power field or more than 1 million round cells per ml. were identified in the semen. Since immature germ cells and white blood cells appear as round cells under wet mount light microscopy, the relationship between increased numbers of round cells in semen and true pyospermia was examined.Semen analyses were performed on specimens from 627 infertility patients. The numbers of round cells per high power field and the number of round cells per ml. of semen were recorded. Specimens from 57 patients (9%) persistently demonstrated greater than 10 round cells per high power field or more than 1 million round cells per ml. and they were subjected to immunohistochemical staining against white blood cell surface antigens. Greater than 1 million white blood cells per ml. were identified in only 20 patients (35%) and only 8 samples demonstrated more than 3 million white blood cells per ml. Of the patients with greater than 1 million white blood cells per ml. 65% had motilities of less than 60% compared to 49% of the patients with less than this amount of pyospermia. This difference was not statistically significant. Round cells counted on a per milliliter basis correlated better with results of immunohistochemical staining than did round cells counted on a per high power field basis. These data demonstrate that the majority of patients with excess round cells in the semen do not have pyospermia and, therefore, empiric antibiotic therapy without further testing will likely be unproductive.

Differentiation of round cells in semen by means of monoclonal antibodies and relationship with male fertility

To differentiate round cells in semen samples of subfertile men and evaluate the clinical significance during infertility investigation. One hundred and eight randomly chosen couples with a median duration of infertility of 4 (range, 1 to 20) years presenting at the outpatient infertility clinic of the University of Heidelberg, Germany. Differentiation of round cells in semen by means of monoclonal antibodies (mABs) and a streptavidin-biotin system for staining. Correlation of results with medical history, outcome of clinical examination, sperm analysis, microbial screening of both partners, evaluation of sperm functional capacity in vivo by means of the postcoital test (PCT) and in vitro with the standardized crossed sperm-cervical mucus penetration test (SCMPT) and the subsequent fertility in a prospective study. The method used for differentiation of round cells proved to be practical and suitable for routine use. The percentage of leukocytes ranged from 0% to 58% with a median of 3%. Number of round cells and percentage of leukocytes did not differ markedly with regard to andrologic history, clinical findings, for example, varicocele, results of standard sperm analysis, and microbial colonization of semen samples. However, high rates of leukocytes of the round cells correlated with reduced sperm count and morphology and results of PCT. Leukocyte-positive (> 15% leukocytes) specimens were also significantly more frequent in case of inadequate SCMPT and reduced sperm penetration ability in vitro. In asymptomatic patients (in terms of genital tract infection), the majority of round cells consist of immature germ cells and < 5% are white blood cells. The streptavidin-biotin system and the mABs used in this study proved to be useful to identify patients with elevated rates of leukocytes in semen possibly reflecting subclinical genital tract infection with influence on sperm functional capacity and subsequent fertility. Thus the procedure can be recommended to be included in a comprehensive evaluation of male fertility.

Ureaplasmal infections of the male urogenital tract, in particular prostatitis, and semen quality.

Ureaplasma urealyticum is considered an etiologic agent in urogenital tract infections, especially prostatitis. Using the 'four-specimen technique', diagnosis can be based upon significant numbers of these microorganisms. In ejaculate, the critical number seems to be 10(3) cfu/ml of semen to discriminate between real infection and contamination during urethral passage. In our study, 46 of 412 samples (11.2%) exceeded this critical number. Most but not all patients suffering from ureaplasma-associated prostatitis established by the 'four-specimen technique' revealed significantly high ejaculate numbers, whereas all samples from patients with prostatodynia and healthy controls had lower numbers. In these cases, numbers of round cells in semen, i.e. all leukocytes and spermatides, were significantly increased as compared to prostatodynia. A significantly negative correlation was detected between the numbers of ureaplasmas and zinc concentration in semen, and an almost identically negative correlation to the content of fructose, thus indicating secretory dysfunction of the accessory glands in ureaplasmal infections of the prostate.