Contamination Of Semen Research Articles

Identification and Antimicrobial Susceptibility Profile of Preputial Bacterial Flora from apparently Healthy Bulls, Rams, and Bucks in Maiduguri, Northeastern Nigeria

The aim of this study was to identify the preputial aerobic bacterial flora from bulls, rams and bucks and to determine their antimicrobial sensitivities. Preputial swabs were collected randomly from twenty each of clinically healthy post pubertal bulls, rams and bucks. Using standard bacteriological techniques, the following bacteria were identified; Escherichia coli (bulls 26.5%, rams 19.2% and buck 22.7%), Staphylococcus aureus (bulls 25.0 %, rams 26.0 % and bucks 24.2 %), Streptococcus (bulls 20.6%, rams 26.0 % and bucks 25.8%), Salmonella (bulls 14.7%), Shigella (rams 15.1% and bucks 15.2%) and Klebsiella (bulls 13.2%, rams 13.7 % and bucks 12.1%) species. There was no statistically significant difference (p>0.05) between the proportions of bacterial species and different animal species. In bulls, all bacterial isolates were susceptible to Ciprofloxacin (100%) and Levofloxacin (100%) while 60% of the bacterial isolates showed resistance to amoxicillin, ampiclox and streptomycin. In rams, all the bacterial isolates were susceptible to ciprofloxacin (100%) and gentamycin (100%) while 40% of bacterial isolates were resistant to chloramphenicol. In bucks, all bacterial isolates were susceptible to amoxicillin (100%), ampiclox (100%) and gentamycin (100%) while 20% bacterial isolates were resistant to streptomycin. This study shows that a few bacteria colonize the prepuce of ruminants in Maiduguri and there is need for preputial washing prior to breeding to reduce contamination of semen during natural mating or collection. Amoxiciillin, ampiclox, ciprofloxacin and gentamycin could be used as therapeutic agents to diseases that could be caused by the bacteria in prepuce of these animal species.

ИЗУЧЕНИЕ БИОЛОГИЧЕСКОЙ ПОЛНОЦЕННОСТИ СПЕРМАТОЗОИДОВ В СПЕРМЕ С БАКТЕРИАЛЬНОЙ ОБСЕМЕНЕННОСТЬЮ

The aim of the study is to investigate the relationship between bacterial contamination of bull semen and the biological value of spermatozoa, as well as reproductive performance. The object of the study was Holstein bulls (n = 17). The material for the study was frozen-thawed semen of bulls in the amount of 61 samples. Microbiological studies of sperm doses were carried out using standard nutrient media in accordance with GOST 32222-2013. The total number of microorganisms was calculated by the number of grown colony-forming units (CFU/ml) in 1 cm3, with subsequent determination of their morphological pro-perties. Sperm motility and morphology were determined using the Argus CASA program. The state of DNA in spermatozoa was studied by the acridine orange test (AO test) using a fluorescence microscope. The study revealed a high positive correlation (r = +0.940**) between the number of microorganisms in sperm and pathology in individual sperm segments. Analysis of variance confirmed a statistically significant effect of the number of microorganisms on sperm morphology F = 123.2 (p=0.000). The correlation relationship between the fragmentation index and the content of microorganisms has a high value (r = +0.965**). Sperm motility negatively correlates (r = –0.768**) with the number of microorganisms (CFU/ml). In the presence of Mycoplasma spp in the samples, the number of cows with fruitful insemination was 35.5 % due to reduced motility, morphology and high fragmentation of sperm DNA (58 %). Sperm motility in samples containing Mycoplasma spp averages 24.5 %, while in samples without these microorganisms it is 48.9 %. Thus, it can be assumed that the presence of Mycoplasma spp in samples and high bacterial contamination rates lead to to lower reproductive performance in bulls.

Prevalence and implications of microplastic contaminants in general human seminal fluid: A Raman spectroscopic study

Microplastics are ubiquitous environmental contaminants that have been detected in human semen from polluted areas, yet their prevalence and effects in the general population remain largely unexplored. To examine microplastic presence, abundance, polymer types, and associations with semen quality parameters in individuals without occupational exposures, this study was conducted by collecting semen samples from 40 participants undergoing premarital health assessments in Jinan, China. Raman microspectroscopy was employed to identify, quantify, and categorize microplastic polymers, sperm motility was assessed via computer-assisted analysis, and morphology was evaluated through Diff-Quik staining. Correlations between demographics, semen parameters, and microplastic content were examined by statistical analysis. We found that microplastics were detected in all semen samples, with 2 particles per sample (ranging from 0.72 to 7.02 μm). Eight distinct polymers were identified, with polystyrene (31 %) being most prevalent. Semen exposed to polystyrene demonstrated higher sperm progressive motility as compared to polyvinyl chloride exposure group (43.52 ± 14.21 % vs 19.04 ± 13.46 %). Sperm morphological abnormalities were observed but not significantly associated with specific plastic types. In conclusion, this study reveals microplastic contamination in semen from individuals without occupational exposure, with PS, PE, and PVC being the most prevalent and exhibiting differential correlations with sperm progressive motility, and highlight the need for further research into the potential reproductive impacts of microplastic exposure.

Comparative Study of Bacterial Contamination in Local Iraqi Sheep and Goats Semen

Background: Semen contamination is a detrimental factor in decreasing fertility. Seasonal changes may affect the contamination, too. Objectives: This study was designed to detect semen contamination in ovine and caprine during different seasons. Methods: Six fully mature male sheep and goats were subjected to electro-ejaculator collection twice monthly from February 1, 2022, to January 31, 2023 (Spring, February 1, 2022-April 30, 2022; Summer, May 1, 2022, July 31, 2022; Autumn August 1, 2022, October 31, 2022; Winter November 1, 2022, January 31, 2023), for studying the seasonal effect. A total of 288 semen samples were collected from both species (36 samples from each per season). All samples were subjected to bacterial isolation and identification. Results: The results indicated that sheep semen had 4 different types of bacterial contamination with a higher number in contaminated samples than goat, which showed 5 different bacterial isolations. There were no significant differences (P>0.05) in bacterial contamination with regard to different seasons. Conclusion: Different seasons appear not to affect bacterial contamination of semen in sheep and goats. Meanwhile, isolating all bacteria types was considered normal flora in both small ruminant species.

Reduced bacterial load in stallion semen by modified single layer centrifugation or sperm washing

The presence of bacteria poses a significant challenge to the quality of stallion semen used in artificial insemination. The bacterial content of insemination doses arises from various sources, such as the healthy stallion, environment, and collection equipment, and is implicated in fertility problems as well as reduced sperm quality during storage. The conventional approach of adding antibiotics to semen extenders raises concerns about antimicrobial resistance and potential negative effects on sperm characteristics, and may not be effective in inhibiting all bacteria. The objective of this study was to determine whether an innovative alternative to antibiotic usage – centrifugation through a single layer of a low density colloid (SLC) – could reduce the bacterial load in stallion semen, and to compare sperm characteristics in samples arising from this procedure, or simple extension of the ejaculate in semen extender, or from sperm washing, i.e. adding extender and then centrifuging the sample to allow the removal of most of the seminal plasma and extender. Eighteen semen samples were collected from six stallions. The semen samples were split and extended prior to washing or SLC, or received no further treatment other than extension. After preparation aliquots from each type of sample were sent for bacteriological examination; the remaining samples were stored for up to 72 h, with daily checks on sperm quality. The low density colloid SLC outperformed sperm washing or extension for bacterial reduction, effectively removing several bacterial species. The bacterial load in the samples was as follows: extended semen, 16 ± 6.7 × 105; washed, 5.8 ± 2.0 × 105; SLC, 2.3 ± 0.88 × 105, p < 0.0001. In addition, SLC completely removed some bacterial species, such as Staphylococcus xylosus. Although there is no selection for robust spermatozoa with the low density colloid, sperm motility, membrane integrity, and DNA fragmentation were not different to washed sperm samples. These findings suggest that SLC with a low density colloid offers a promising method for reducing bacterial contamination in stallion semen without resorting to antibiotics.

Strategies for Bacterial Eradication from Human and Animal Semen Samples: Current Options and Future Alternatives.

The primary role of semen processing and preservation is to maintain a high proportion of structurally and functionally competent and mature spermatozoa, that may be used for the purposes of artificial reproduction when needed, whilst minimizing any potential causes of sperm deterioration during ex vivo semen handling. Out of a multitude of variables determining the success of sperm preservation, bacterial contamination has been acknowledged with an increased interest because of its often unpredictable and complex effects on semen quality. Whilst antibiotics are usually the most straight-forward option to prevent the bacterial contamination of semen, antimicrobial resistance has become a serious threat requiring widespread attention. As such, besides discussing the consequences of bacteriospermia on the sperm vitality and the risks of antibiotic overuse in andrology, this paper summarizes the currently available evidence on alternative strategies to prevent bacterial contamination of semen prior to, during, and following sperm processing, selection, and preservation. Alternative antibacterial supplements are reviewed, and emphasis is given to modern methods of sperm selection that may be combined by the physical removal of bacteria prior to sperm preservation or by use in assisted reproductive technologies.

Short-Term Storage of Rooster Ejaculates: Sperm Quality and Bacterial Profile Differences in Selected Commercial Extenders.

Bacterial contamination of semen has become an important contributor to the reduced shelf life of insemination doses in the poultry industry, which is why antibiotics (ATBs) are an important component of semen extenders. Due to a global rise in antimicrobial resistance, the aim of this study was to assess the efficiency of selected commercially available semen extenders to prevent possible bacterial contamination of rooster ejaculates. Two selected extenders free from or containing 31.2 µg/mL kanamycin (KAN) were used to process semen samples from 63 healthy Lohmann Brown roosters. Phosphate-buffered saline without ATBs was used as a control. The extended samples were stored at 4 °C for 24 h. Sperm motility, viability, mitochondrial activity, DNA integrity and the oxidative profile of each extended sample were assessed following 2 h and 24 h of storage. Furthermore, selective media were used to quantify the bacterial load and specific bacterial species were identified with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The results indicate that semen extenders enriched with KAN ensured a significantly higher preservation of sperm quality in comparison to their KAN-free counterparts. Bacterial load was significantly decreased in diluents supplemented with ATBs (p ≤ 0.001); however, KAN alone was not effective enough to eradicate all bacteria since several Escherichia coli, Enterococcus faecalis, Enterococcus faecium and Micrococcus luteus were retrieved from samples extended in KAN-supplemented commercial extenders. As such, we may suggest that more focus should be devoted to the selection of an optimal combination and dose of antibiotics for poultry extenders, which should be accompanied by a more frequent bacteriological screening of native as well as extended poultry semen.

P-167 Bacterial contamination of embryo culture medium in conventional in vitro fertilisation versus ICSI

Abstract Study question Our aim is presenting a clinical case of pregnancy using intracytoplasmatic sperm injection with bacterial contamination of embryo culture after failed conventional in vitro fertilisation. Summary answer Intracytoplasmatic sperm injection (ICSI) can be an effective and safe method to prevent embryo culture infection in case of microbial contamination of semen. What is known already Contamination of embryo culture is rare, but can be detrimental. The most common sources are semen and follicular fluid. We currently have no standard protocols for monitoring biological fluids. Semen often contains bacteria even if hygiene rules for collection are followed. Most studies found no significant difference in the effectiveness of in vitro fertilisation treatment in the presence or absence of bacteriospermia. This can be explained by sperm preparation techniques, antibiotic content of culture media and use of ICSI. Most centres advise against contaminated embryo transfers, but some achieved pregnancy after removing the zona pellucida or using ICSI. Study design, size, duration This is a case report of two consecutive in vitro fertilisation - ICSI cycles at the Division of Assisted Reproduction, Department of Obstetrics and Gynecology, Semmelweis University, Budapest in 2022. Participants/materials, setting, methods First we performed conventional in vitro fertilisation in a couple with secondary infertility because of oligoasthenozoospermia. For the second time we suggested ICSI due to bacterial growth observed in the embryo culture medium. Microbiological analyses were carried out. We video-recorded the cytoplasmic infection of one oocyte following ICSI with time-lapse microscopy. Main results and the role of chance After conventional in vitro fertilisation we observed a proliferation of rods in the embryo culture medium. The control culture medium was sterile. The semen and infected embryo culture medium contained Escherichia coli and Staphylococcus hominis. The oocytes did not fertilize and degenerated. The husband was asymptomatic and did not have alteration in physical examination therefore he was not treated with antibiotics. In next cycle we performed ICSI. 2 of 14 oocytes fertilized normally and developed into high-quality blastocysts. One embryo was transferred resulting in live birth and one embryo was cryopreserved. We video-documented proliferation of bacteria in one fertilised oocyte but not in the others. Infected oocyte was removed from culture and its culture medium showed Escherichia coli and Cutibacterium acnes. Fungal culture and sexually transmitted diseases were negative. The semen only contained Escherichia coli. The cervix culture was negative. Escherichia coli was resistant to gentamicin contained in culture medium. Limitations, reasons for caution We only had one case of bacterial contamination in 30 years and therefore can not conclude to the routine use of ICSI if microbial infections of biological fluids are present. Wider implications of the findings ICSI can be offered to patients with bacterial contamination of semen and can help reducing the risk of infection of embryo culture. We can prevent the disadvantages of excessive antibiotic treatments. New guidelines are needed how to handle bacterial contamination of embryo culture media. Trial registration number not applicable

Double semen collection at a 1-h interval in dogs decreases the bacterial contamination of canine ejaculates

Semen extenders usually contain antibiotics with the aim to minimize bacterial growth, but the indiscriminate use of antibiotics increases the emergence of multidrug-resistant bacteria. A limiting factor of semen processing in dogs is the low total sperm count that limits the number of insemination doses that can be obtained from one ejaculate. Therefore, two ejaculates collected at a short interval can be combined to increase the number of AI doses. In this study, semen was collected from dogs either once or the same dogs (n = 28) were submitted to dual semen collection 1 h apart. All ejaculates were submitted to bacteriological analysis. We hypothesized that bacterial contamination of semen is low but that a dual semen collection might increase contamination. A sample for bacteriological examination was taken from raw semen immediately after semen collection. Bacteria including mycoplasmas were isolated using conventional cultivation procedures and isolates were identified to the species level by matrix-assisted laser desorption ionization – time of flight (MALDI-ToF) mass spectrometry. In total, 22 bacterial species were identified in the 84 ejaculates with Mycoplasma cynos, Streptococcus canis and Canicola haemoglobinophilus being most frequent. Bacterial growth was sporadic in 16 and absent in 10 ejaculates. The overall bacterial growth was lower in the second than in the first ejaculate of dual semen collections (p < 0.05). The percentage of motile and membrane-intact spermatozoa in frozen-thawed ejaculates was not associated with the degree of bacterial contamination of raw semen. In conclusion, there was only limited microbial contamination in dog semen and the microorganisms isolated are considered part of the normal genital bacterial flora. Repeated semen collection reduced bacterial contamination in the second in comparison to the first ejaculate. The use of antibiotics in canine semen should be questioned.

Lohmann Brown Rooster Semen: Intrinsic Bacteria and Their Impact on Sperm Progressive Motility and Seminal Biochemical Parameters—A Preliminary Study

Semen quality plays a crucial role in poultry production; however, it may be impaired by the presence of numerous bacterial species. This study researched the impact of bacterial contamination of Lohmann brown rooster semen on the biochemical parameters of seminal plasma to evaluate its potential consequences on the sperm progressive motility. Semen was collected from 27 stud roosters, and the sperm concentration and progressive motility were measured using computer-assisted semen analysis (CASA). Seminal plasma was separated, and selected biochemical parameters were measured using commercially available assays. An aliquot of each semen sample was cultured, the colonies were counted and the MALDI Biotyper was used for bacterial identification. The samples were divided into three categories based on their sperm progressive motility and the data were compared and statistically evaluated. Moreover, Pearson’s correlation analysis was performed. The results showed that the lower the sperm progressive motility, the higher the level of colony-forming units. Moreover, sperm concentration was significantly higher (p &lt; 0.05) in the group with the highest bacterial occurrence and the lowest proportion of progressively motile spermatozoa. Calcium, magnesium, creatinine, uric acid, alkaline phosphatase, and total proteins significantly changed in semen samples with the lowest proportion of progressive motility. In conclusion, seminal plasma biochemistry may mirror changes occurring in semen as a result of bacterial presence in the reproductive tract of poultry.

Analysis of the clinical outcomes of microbial contamination caused by environmental contamination of the embryology laboratory during IVF-ET treatment cycles

BackgroundBacterial contamination may cause loss of or damage to cultured oocytes or embryos, resulting in the lack of transplantable embryos during IVF embryo culture. However, there are few reports about IVF embryo contamination caused by embryology laboratories. In this work, we evaluated clinical pregnancy outcomes and the risk of maternal and infant complications after embryo contamination caused by environmental pollution during IVF.MethodsThe authors retrospectively analyzed 2490 IVF-ET ovulation induction therapy cycles in the Reproductive Center of Yichang Central People's Hospital from January 2015 to May 2022. According to the presence or absence of embryo culture medium contamination, the two groups were divided into an embryo contamination cycle and a nonembryo contamination cycle. The primary outcome parameters were the characteristics and progress of embryo culture medium contamination. Embryo laboratory outcomes, pregnancy outcomes, and maternal and infant complications were secondary outcome parameters.ResultsOne case of embryo contamination originated from semen contamination. The remaining 15 cases involved environmental contamination outbreaks in embryo culture chambers, caused by Staphylococcus pasteuri. Compared with conventional uncontaminated IVF cycles, the 15 cases of contaminated embryo cycles showed no significant difference in embryo laboratory outcomes, pregnancy outcomes, or maternal and infant complications except for a slightly higher rate of fetal growth retardation. Ultimately, 11 live-born infants were successfully delivered, of which 2 were premature. The remaining 4 patients did not become pregnant after 1–2 transfers due to a lack of transferable embryos.ConclusionWhen the embryo culture medium is contaminated due to the environmental contamination of the IVF culture room, it is feasible to perform daily rapid rinsing of the culture medium and avoid blastocyst culture as remedial treatment. However, the long-term impact on offspring needs further prospective research.

Volatiles from Pseudomonas palleroniana Strain B-BH16-1 Suppress Aflatoxin Production and Growth of Aspergillus flavus on Coix lacryma-jobi during Storage

Semen coicis is not only a traditional Chinese medicine (TCM), but also a typical food in China, with significant medical and healthcare value. Because semen coicis is rich in starch and oil, it can be easily contaminated with Aspergillus flavus and its aflatoxins (AFs). Preventing and controlling the contamination of semen coicis with Aspergillus flavus and its aflatoxins is vital to ensuring its safety as a drug and as a food. In this study, the endosphere bacteria Pseudomonas&nbsp;palleroniana strain B-BH16-1 produced volatiles that strongly inhibited the mycelial growth and spore formation activity of A. flavus. Gas chromatography-mass spectrometry profiling revealed three volatiles emitted from B-BH16-1, of which 1-undecene was the most abundant. We obtained authentic reference standards for these three volatiles; these significantly reduced mycelial growth and sporulation in Aspergillus, with dimethyl disulfide showing the most robust inhibitory activity. Strain B-BH16-1 was able to completely inhibit the biosynthesis of aflatoxins in semen coicis samples during storage by emitting volatile bioactive components. The microscope revealed severely damaged mycelia and a complete lack of sporulation. This newly identified plant endophyte bacterium was able to strongly inhibit the sporulation and growth of Aspergillus and the synthesis of associated mycotoxins, thus not only providing valuable information regarding an efficient potential strategy for the prevention of A. flavus contamination in TCM and food, but potentially also serving as a reference in the control of toxic fungi.

Bacteriospermia among smallholder artificial insemination boars in the Philippines and potential associated factors

Objective: To determine the prevalence of bacteriospermia, the bacterial load, and the potential factors associated with bacterial contamination in boar semen collected by local smallholder artificial insemination operators. Methods: Fifteen individual raw semen samples were collected from locally available artificial insemination boars owned by different smallholder boar operators within the 5th district of Leyte, Philippines and were subjected to standard bacteriological culture and identification, including a survey of potentially associated factors. Prevalence and bacterial count were determined accordingly, while boar characteristics and collection practices were clustered following agglomerative hierarchical clustering technique. Results: One hundred percent contamination with a bacterial count of (2.01±0.38)×103 CFU/mL was observed. At least 73.33% of the samples were positive for Bacillus spp., while other identified isolates included Enterobacter spp., Staphylococcus spp., E. coli, Pseudomonas spp., Citrobacter spp., and Klebsiella spp. Conclusions: Despite the high prevalence of bacteriospermia, the bacterial count is low. Nevertheless, on-farm practices on boar health and management, semen collection, and sanitation as well as the enhancement of basic protocols to control contamination should be conscientiously considered in smallholder artificial insemination operation.

Candida Genus Maximum Incidence in Boar Semen Even after Preservation, Is It Not a Risk for AI though?

There is little information in the literature about the fungal contamination of boar semen and its persistence during storage. The challenge of this study was to perform a mycological screening to identify the yeast in the raw semen at 12/24 h after dilution. The research was done in pig farms in the N-E area of Romania, with maximum biosecurity and state-of-the-art technology. All the examined ejaculates (101) were considered to be normal for each spermogram parameter, with microbiological determinations in T0 at the time of ejaculate collection, T1 at the time of dilution, and T2 at 24 h of storage. Microbiological determinations (mycological spermogram) were performed for quantitative (LogCFU/mL) and qualitative (typification of fungal genera) identification. Bacterial burden (×103 LogCFU/mL) after dilution (T1) decreased drastically (p &lt; 0.0001) compared to the one in the raw semen (T0). After 24 h of storage at 17 °C, the mean value of the bacteriospermia remained constant at an average value of 0.44. Mycospermia had a constant trend at T0 (raw) and T1 (0.149 vs. 0.140) and was slightly higher at T2 (0.236). The difference between T1 vs. T2 (p = 0.0419) was close to the statistical reference value (p = 0.05). Of the total genera identified (24), the fungi had a proportion of 37.4% (9/15) and a ratio of 1:1.6. Regarding the total species (34), the fungi had a frequency of 29.42% (10/24) with a ratio between the fungi and bacteria of 1:2.4. A fertility rate of 86% was observed in the L1 group (50 AI sows with doses and mycospermia from T1), and an 82% rate was observed in the L2 group (50 AI sows with doses and mycospermia from T2). The litter size of L1 was 9.63 piglets and 9.56 for L2. Regarding the total number of piglets obtained between the two groups, there was a slight decrease of 22 piglets in group L2, without statistical differences (p &gt; 0.05). The predominant genera persisted after dilution during a 12 h storage at 17 °C, where yeasts, such as Candida parapsilosis and C. sake were identified in more than 92% of AI doses.

Screening for Taylorella equigenitalis in Equine Semen: An Exploratory Study

The study examined and compared the sensitivity of culture and a quantitative PCR assay for screening equine semen for the presence of Taylorella equigenitalis (CEMO). Chilled semen samples, both raw and treated with extender, from two stallions were spiked with the organism at seven or 23 days postejaculation and prepared in serial dilutions. Culture of the 7-day raw semen readily detected CEMO at all dilutions, but extended semen yielded counts that were two log cycles lower at equivalent dilutions, with the organism being nearly undetectable at the maximal dilutions. By contrast, PCR sensitivity was not affected by extender, but for 7-day-old raw semen, PCR detection declined abruptly three log dilutions earlier than detection by culture. The more aged 23-day-old semen proved less satisfactory for spiking, with detection of CEMO by culture failing in three of the four samples due to overgrowth with commensal organisms. However, PCR performance was similar in both the 23- and 7-day spiking series. The detection limit by PCR is estimated at between 104 and 105 cfu/mL. Typical CEMO concentrations in the semen of colonized stallions are not widely reported but where natural semen contamination has been investigated, the organism was present at this order of magnitude. The reliability of detecting CEMO infection using semen samples by either method is discussed.

To study the effect of microbial contamination in semen and its effect on male infertility

According to the 3rd WHO methods, normal sperm morphology has been considered to be directly related with the fertilization process and pregnancy. The quality of semen such as sperm morphology, count, and motility has been reduced due to abnormalities in male thus, causing infertility. A considerable factor responsible for infertility among male partners may be as a result of microbial contamination. Semen may contain contamination while collecting sample. However, there are some sperm preparation techniques that can be used for the removal of contamination in semen such as; swim up, density gradient, simple wash, discontinuous density gradient. The main objective of this study is to find out the presence of infections and microbial contamination. This study will compare sperm count and sperm motility of patient’s semen sample with numbers of debris/ present infectious agent. Microbiological assay of semen sample will be performed. Also, various infectious agents causing infertility will be evaluated and studied. Semen analysis, semen preparation by density gradient, semen culture were carried out to look into the effect of microbial contamination on semen sample of infertile men.

Sexually transmitted doping: The impact of urine contamination of semen.

The high sensitivity of antidoping detection tests creates the possibility of inadvertent doping due to an athlete's unknowing ingestion of contaminated environmental sources such as dietary supplements, food, or drinks. Recently, athletes denying use of a prohibited substance have claimed that the positive antidoping tests was due to exchange of bodily fluids with a nonathlete partner using a prohibited substance. Measurement of drugs in semen is largely limited to one or very few samples due to the inaccessibility of sufficiently frequent semen samples for detailed pharmacokinetics. An emerging issue in semen drug measurements is that semen samples may contain residual urine from ejaculation left in the urethra; however, the urine content in semen samples has not been studied. In the present study, we employed concurrent creatinine measurements in urine and seminal plasma to determine the urine content of semen samples.

The Impact of Bacteriocenoses on Sperm Vitality, Immunological and Oxidative Characteristics of Ram Ejaculates: Does the Breed Play a Role?

Simple SummaryRam semen frequently presents with bacterial contamination, which may affect the resulting sperm vitality and fertilization ability. Since semen quality is of paramount importance for the successful artificial insemination of ewes, we focused on the description of bacterial profiles of ejaculates collected from three Slovak sheep breeds. Furthermore, we strived to unravel the effect of bacteriospermia on the immunological characteristics and oxidative profile of semen, both of which seem to play a role in bacteria-inflicted damage to male gametes.Bacterial contamination of semen is an often overlooked, yet important, factor contributing to decreased sperm vitality. Understanding the impact of bacterial presence on sperm structural integrity and functional activity may assist the development of effective strategies to prevent, or manage, bacteriospermia in the breeding practice. The aim of this study was to describe the bacterial profiles of ram semen (n = 35), and we also focused on the associations between bacteriospermia, sperm structure, and function, as well as oxidative and inflammatory characteristics of semen. For a better insight, the samples were divided into three groups, according to the breeds used in the study: native Wallachian (NW), improved Wallachian (IW), and Slovak dairy (SD) breeds. The results showed a significantly lower motility and membrane integrity in the NW group in comparison to the IW and SD groups, which was accompanied by a significantly higher concentration of leukocytes, increased reactive oxygen species (ROS) generation, and subsequent oxidative insults to the sperm lipids and proteins. Accordingly, the NW group presented with the highest bacterial load, in which Staphylococcus and Escherichia were the predominant representatives. The Pearson correlation analysis uncovered positive relationships amongst the bacterial load and leukocytospermia (r = 0.613), the extent of lipid peroxidation (r = 0.598), protein oxidation (r = 0.514), and DNA fragmentation (r = 0.638). Furthermore, positive correlations were found between the bacterial load and pro-inflammatory molecules, such as the C-reactive protein (r = 0.592), interleukin 1 (r = 0.709), and interleukin 6 (r = 0.474), indicating a possible involvement of the immune response in the process of bacteriospermia. Overall, our data indicate that ram semen quality may be equally affected by the bacterial load and diversity. Furthermore, we can assume that the presence of bacteria in ejaculates triggers inflammatory processes, causes ROS overproduction, and, thereby, contributes to alterations in the sperm structure, while at the same time compromising the fertilization ability of male gametes.

Boar Semen Contamination: Identification of Gram-Negative Bacteria and Antimicrobial Resistance Profile

Simple SummaryBoar semen can contain many bacterial species, some of which can have a negative impact upon the quality of the semen, as well as on the sows’ reproductive capacity. Semen contamination may occur at time of collection or during semen processing. The aim of this study was to identify gram-negative bacteria that appear in boar semen and to establish models of antimicrobial resistance of isolated gram-negative bacteria. Semen doses examined contained bacterial species with a known negative effect on sows’ reproductive tracts (Pseudomonas, Enterobacter, Klebsiella, E. coli), and more than half of these isolates were resistance to gentamycin (56.52%) and penicillin (58.69%) antimicrobials commonly used in boar semen extenders. This work proved the presence of pathogenic multiple resistant bacteria in semen, and therefore, we highly recommend periodic microbiological screening of bacteriospermia in boars to avoid the use of low-quality semen in the pig industry.Bacterial contamination of boar semen occurs with some frequency in artificial insemination centers and may have a negative effect on the quality of the semen as well as on the sows’ reproductive capacity. Normally, the source of bacterial contamination in pig seminal doses is the own boar. However, distilled water or laboratory equipment used to elaborate the seminal doses can be an important source of bacterial contamination. This study focused on the identification of gram-negative bacteria in boar semen, and impact on the quality of ejaculates obtained from boar, as well as on the establishment of antimicrobial resistance patterns of isolated gram-negative bacteria. Semen samples were collected from 96 boars, ranging in age from 12–36 month, from three artificial insemination centers from the North-West of Romania. Bacterial species were identified by two methods: matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and API 20 E (BioMerieux, France). The main bacteria isolated from the doses diluted semen were gram-negative bacteria (47.91%), with a majority of the contaminant bacteria belonging to the family Enterobacteriaceae: Seratia marcescens 19.56%, Proteus mirabilis 15.21% and Escherichia coli 10.86% and to the family Pseudomonaceae: Ralstonia picketii 17.39%, Burkholderia cepacia 10.86%, Pseudomonas aeruginosa 8.69%, and Pseudomonas fluorescens 4.34%, respectively. More than half of these isolates (56.52%) were resistant to gentamycin and 58.69% were resistant to penicillin. These antibiotics are very frequently added in sperm diluent in the centers for the processing of sperm from boars in Romania. Regular monitoring for bacterial contamination is an important aspect of a control program.

Identification of Bull Semen Microbiome by 16S Sequencing and Possible Relationships with Fertility.

Reports on the use of 16S sequencing for the identification of bacteria in healthy animals are lacking. Bacterial contamination of bull semen can have a negative effect on the sperm quality. The aims of this study were threefold: to identify bacteria in the semen of healthy bulls using 16S sequencing; to investigate the differences in the bacterial community between individual bulls; and to establish if there was a relationship between the bacteria isolated and bull fertility. Semen from 18 bulls of known fertility was used for the DNA extraction and 16S sequencing; 107 bacterial genera were identified. The differences in the amplicon sequence variants (ASVs) and the numbers of genera between bulls were noted. Negative correlations (p < 0.05) between several bacterial genera with Curvibacter, Rikenellaceae RC9-gut-group and Dyella spp. were seen. Other negatively correlated bacteria were Cutibacterium, Ruminococcaceae UCG-005, Ruminococcaceae UCG-010 and Staphylococcus, all within the top 20 genera. Two genera, W5053 and Lawsonella, were enriched in bulls of low fertility; this is the first time that these bacteria have been reported in bull semen samples. The majority of the bacteria were environmental organisms or were species originating from the mucous membranes of animals and humans. The results of this study indicate that differences in the seminal microbiota of healthy bulls occur and might be correlated with fertility.