1. SEMEN ANALYSIS AND SPERM CULTIVATION
In order for a spermogram to be reliable, it should be done after abstinence from sexual intercourse for 3-4 days, which does not include the day of last intercourse and the day of semen collection. It is still necessary to have proper arousal before taking a sample, which is why it is often necessary for his partner to be present and participate in the procedure if she wishes. Before collecting the sperm, the genital area and hands should be thoroughly washed. During ejaculation, through masturbation, the sperm is collected in a sterile container, which you can either buy from the pharmacy (it is similar to the urine collector), or we will offer you one at Medimall IVF Clinic.
The collection of sperm takes place in our private, especially designed space. If the semen is obtained at home, be careful when transporting the sperm sample to the laboratory, to maintain the body temperature. This can be achieved by placing it in contact with the body or by wrapping the container with cotton and aluminum foil externally.
From the moment the sperm is collected, until the moment it is delivered to the laboratory, it should not take more than an hour. If the semen culture shows that a germ is present in the semen, treatment with the appropriate antibiotics, selected from the antibiogram along with the microbe-positive semen culture, is required.
As for the values of the sperm parameters, if they are lower than normal, the semen analysis should be repeated at least 15 days from the previous. If the second sperm chart shows that the parameters of the sperm are lower than normal (defined by the World Health Organization), the sperm is characterized as abnormal and further testing is recommended depending on its pathology. The additional tests that the partner should undergo depending on the case are:
- examination by a urologist andrologist
- scrotum doppler
- hormonal control
- test for cystic fibrosis
- sperm DNA fragmentation
2. EXAMINATION BY UROLOGIST ANDROLOGIST
Clinical examination by a specialized andrologist or urologist to detect all those pathological conditions that could result a pathological sperm count respectively, such as hypoplastic or atrophic testicles, cryptorchidism, etc.
3. DOPPLER SCROTUM
It is a non-invasive test used to assess the testicles, epididymis, and blood flow to the testicles. You do not need any special preparation for the examination, while after the ultrasound, you can continue your daily activities without any restrictions. In case the examination indicates some kind of pathology, the doctor will give you all information needed, additional instructions, and appropriate personalized treatment options.
4. HORMONAL CONTROL
When controlling a man’s infertility, it is necessary to measure the levels of specific hormones that could directly or indirectly affect sperm production. The main hormones that should be checked are testosterone, FSH, LH, prolactin, TSH, E2, and cortisol.
Examining the hormonal profile of a man, it is important to know the reasons, why the man there is has a reduced sperm count, e.g. varicose veins, testicular cancer, obstruction of the seminal vesicles, etc. Reduced testicular size or underdeveloped secondary sexual characteristics in men (penis size, pubic hair, reduced muscle mass, lack of body hair, etc.) also require hormonal investigation.
5. SPERM DNA FRAGMENTATION
Sperm DNA fragmentation is now one of the leading causes of male infertility. The simple parameters of the sperm diagram – concentration, motility, morphology –cannot detect fragmentation. In addition, samples with normal parameters are very likely to show high fragmentation rates. The high degree of DNA fragmentation could cause such suspension to a successful pregnancy, to the point that the fertility treatment followed by the partner does not matter anymore. Several studies have shown that increased levels of DNA fragmentation are associated with reduced pregnancy and birth rates, poor fetal quality, and increased miscarriage.
High levels of DNA fragmentation in sperm could also be caused by factors such as:
- exposure to environmental pollutants
- your profession (eg driver of vehicles)
- advanced age
- malnutrition and obesity
- increase in testicular temperature (eg from laptops)
- drug use
- high fever
- the varicose vein
- cancer and its treatments
The test requires a small amount of sperm and after appropriate processing in the laboratory, the sperms with fragmented DNA are detected, which acquire a different morphology from those with whole DNA. The test results are expressed as DNA Fragmentation Index -% DFI (percentage of sperm with intact DNA). The test results are indicated as DNA Fragmentation Index -% DFI (percentage of sperm with fragmented DNA).
Depending on their percentage, they are divided into the following categories:
- <15% DFI high potential fertility
- > 15% to <30% DFI good potential fertility> 30% DFI low potential fertility
The result is defined as normal when the DFI fraction (%) in the sperm is below 30%. Normal pregnancies occur even in couples where the husband has a high rate of DNA fragmentation in the sperm, but still with a reduced chance of pregnancy and an increased risk of miscarriage.
Is there a cure for sperm DNA fragmentation? A series of “healthy” habits and methods could contribute significantly for infertile men to enhancing fertility, by reducing their sperm fragmentation. Who are they:
- smoking cessation
- a high-antioxidant diet, given that oxidative stress is the main cause of DNA fragmentation
- avoidance of certain drugs
- checking for any urinary tract infections
- healthy eating
- proper information from your doctor about the vitamins and antioxidant supplements suitable for boosting fertility.
6. MOLECULAR DETECTION (PCR) OF CHLAMYDIA, UREAPLASMA, MYCOPLASMA
Sexually transmitted infections such as chlamydia, ureaplasma and mycoplasma are the most common. Chlamydia and more specifically chlamydia trachomatis, is the most common sexually transmitted infection. Mycoplasma (mainly mycoplasma hominis) and ureaplasma (mainly ureaplasma urealyticum), are bacteria that are often detected in the female genitals and are transmitted through sexual contact. Their existence in the body could reduce fertility. Their presence could be detected by semen culture.
7. KARYOTYPE BLOOD EXAMINATION
The representation of the set of chromosomes of an individual is called a karyotype. The normal human karyotype contains 46 chromosomes (23 pairs), including the two sex-determining chromosomes. The karyotype of peripheral blood lymphocytes enables the control of all chromosomes for the possible existence of numerical and structural chromosomal abnormalities and rearrangements. As cytogenetic abnormalities are associated with both male and female infertility, the classic karyotype concerns couples with infertility problems, with two or more first-trimester miscarriages and previous failed IVF attempts. The detection of such abnormalities is crucial for the proper planning of a subsequent pregnancy that can occur either spontaneously or by assisted reproduction methods. Peripheral blood karyotype must be submitted to persons/couples who had in their family history:
a. People with:
- chromosomal abnormalities
- mental retardation of unknown cause
- multiple miscarriages and neonatal death
- primary or secondary amenorrhea
- premature menopause
- sperm abnormalities (azoospermia, oligospermia)
- abnormal clinical phenotype (delayed or excessive physical development, deformity, mental retardation)
b. Couples with:
- infertility of unknown cause (inability to conceive naturally or through assisted reproduction procedures)
- multiple miscarriages or a history of undetected neonatal death
- child with a diagnosed chromosomal abnormality
- diagnosed chromosomal abnormality of a structural nature in the karyotype of fetal amniotic fluid during pregnancy
8. CHECK FOR CYSTIC FIBROSIS
Cystic fibrosis is the second most common genetic disease in Greece, after Mediterranean anemia, and the first in the Caucasian race. The disease is due to mutations in the CFTR (cystic fibrosis transmembrane conductance regulator) gene, and is inherited in its residual nature. This means that the carriers of the mutated gene are perfectly healthy, but have a 50% chance of passing on their mutated gene to their offspring.
The disease has a wide range of symptoms both in severity and extent (respiratory, digestive and other systems). This means that there are variants in the CFTR gene that are considered mild and do not bring about the characteristic pathological phenotype of the disease, they are simply related to hypersensitivity of the respiratory system (eg frequent respiratory infections). In addition, men with cystic fibrosis are sterile and 20-40% are often carriers of a single abnormal CFTR gene and are therefore considered clinically healthy.
With the current data in our country, the screening test should be applied to all high-risk groups, but also to all couples who are going to have a child, at the level of prenatal control. In the latter case, the testing starts from the mother, to investigate whether she is a carrier of a mutation in the fibrocystic disease. If the mother is found to be a carrier of such a mutation, then it is necessary to examine the father with extended mutation control, since the risk of giving birth to a child with the disease increases greatly. It should be noted that many couples with male infertility should undergo genetic testing for cystic fibrosis-related mutations at the same time: a significant proportion of men with obstructive azoospermia undergo micro fertilization techniques. carriers of mutations in the CFTR gene.
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