Seminal plasma inhibits Chlamydia trachomatis infection in vitro, and may have consequences on mucosal immunity.

Seminal plasma (SP) is the main vector of C. trachomatis (CT) during heterosexual transmission from male to female. It has immunomodulatory properties and impacts the susceptibility to HIV-1 infection, but its role has not been explored during CT infection. In the female reproductive tract (FRT), CT infection induces cytokine production and neutrophil recruitment. The role of neutrophils during CT infection is partially described, they could be at the origin of the pathology observed during CT infection. During this study, we developed an experimental in vitro model to characterize the impact of CT infection and SP on endocervical epithelial cell immune response in the FRT. We also studied the impact of the epithelial cell response on neutrophil phenotype and functions. We showed that the production by epithelial cells of pro-inflammatory cytokines increased during CT infection. Moreover, the pool of SP as well as individuals SP inhibited CT infection in a dose-dependent manner. The pool of SP inhibited cytokine production in a dose-dependent manner. The pool of SP altered gene expression profiles of infected cells. The culture supernatants of cells infected or not with CT, in presence or not of the pool of SP, had an impact on neutrophil phenotype and functions: they affected markers of neutrophil maturation, activation and adhesion capacity, as well as the survival, ROS production and phagocytosis ability. This study proposes a novel approach to study the impact of the environment on the phenotype and functions of neutrophils in the FRT. It highlights the impact of the factors of the FRT environment, in particular SP and CT infection, on the mucosal inflammation and the need to take into account the SP component while studying sexually transmitted infections during heterosexual transmission from male to female.

Male Tract Microbiota and Male Infertility.

In recent years, several studies have analyzed the composition of the male genital tract microbiota and its changes in infertility or in different situations associated with infertility. The aim of this narrative review is to obtain more insight on this topic; in particular, to describe actual evidence about changes in the semen microbiota in patients with infertility, male tract infections, or HPV infections. In semen, an increase in semen Prevotella spp. is associated with oligozoospermia and with obesity-associated asthenozoospermia; an increase in Pseudomonas is more frequently associated with asthenozoospermia and oligozoospermia; a reduction in Lactobacilli spp. (namely in Lactobacillus crispatus) may represent a marker of low semen quality. However, an increase in Lactobacillus iners is considered a risk factor for a reduced sperm concentration. In patients with prostatitis, there is a reduction in Lactobacillus spp. and an increase in Streptococcus spp., opening important perspectives about the role of probiotic treatments in these patients. Finally, an increase in Fusobacteria spp. was observed in patients with an HPV infection. In the conclusion, we underline the interactions between the seminal and vaginal microbiota, so that further studies should focus on the "couple genital microbiota".

Low-density colloid centrifugation removes bacteria from boar semen doses after spiking with selected species.

Single-layer centrifugation (SLC) with a low-density colloid is an efficient method for removing contaminating microorganisms from boar semen while recovering most spermatozoa from the original sample. This study tested the performance of this technique, using 50-ml tubes, by spiking commercial semen doses prepared without antibiotics with selected bacterial species followed by storage at 17 °C. The doses were spiked up to 102/ml CFU (colony forming units) of the bacteria Burkholderia ambifaria, Pseudomonas aeruginosa, and Staphylococcus simulans. The semen was processed by SLC (15 ml of sample and 15 ml of colloid) with the colloid Porcicoll at 20% (P20) and 30% (P30), with a spiked control (CTL) and an unspiked control (CTL0), analyzing microbiology and sperm quality on days 0, 3 and 7. SLC completely removed B. ambifaria and S. simulans, considerably reducing P. aeruginosa and overall contamination (especially P30, ∼104 CFU/ml of total contamination on day 7, median). Sperm viability was lower in P20 and P30 samples at day 0, with higher cytoplasmic ROS. Still, results were similar in all groups on day 3 and reversed on day 7, indicating a protective effect of SLC (possibly directly by removal of damaged sperm and indirectly because of lower bacterial contamination). Sperm chromatin was affected by the treatment (lower DNA fragmentation and chromatin decondensation) and storage (higher overall condensation on day 7 as per chromomycin A3 and monobromobimane staining). In conclusion, SLC with low-density colloids can remove most bacteria in a controlled contamination design while potentially improving sperm quality and long-term storage at practical temperatures.

Evaluation and management of male genital tract infections in the setting of male infertility: an updated review.

PURPOSE OF REVIEW: Male infertility may be secondary to male genital tract infection (MGTI) in an estimated 15% of cases. In the absence of overt clinical signs, evaluation for MGTI beyond semen analysis is not well established. Therefore, we review the literature on the evaluation and management of MGTI in the setting of male infertility. RECENT FINDINGS: A set of international guidelines recommends semen culture and PCR testing, but the significance of positive results remains unclear. Clinical trials evaluating anti-inflammatory or antibiotic treatment report improvements in sperm parameters and leukocytospermia, but data on the effect on conception rates are lacking. Human papillomavirus (HPV) and the novel coronavirus (SARS-CoV-2) have been associated with poor semen parameters and decreased conception rates. SUMMARY: The finding of leukocytospermia on semen analysis prompts further evaluation for MGTI, including focused physical examination. The role of routine semen culture is controversial. Treatment options include anti-inflammatories; frequent ejaculation; and antibiotics, which should not be used in the absence of symptoms or microbiological infection. SARS-CoV-2 represents a subacute threat to fertility that should be screened for in the reproductive history along with HPV and other viruses.

Effects of air exposure and agitation on quality of stored boar semen samples.

The objective of this study was to investigate the effects of semen volume, air contact inside semen dose tubes, daily agitation of semen doses and extender type on semen quality, thermo-resistance and bacteria growth in extended boar semen doses preserved over 7 days of liquid storage. Ejaculates from 4 proven terminal cross-bred boars were collected using the gloved-hand technique for 4 weeks and used in the 3 × 2 × 2 factorial study. The effects of treatment (CON: 80 ml doses sealed at the top of the tube; 40HIGH: 40 ml doses sealed at top of tube, and 40LOW: 40 ml doses sealed at top of the liquid), agitation (agitated versus not agitated) and extender type (long-term versus short-term) were investigated on semen quality, thermo-resistance and bacteria growth in boar semen doses. The results of the study revealed that motility (p = .031) and viability (p = .041) in 40HIGH were lower than CON. pH (p < .001) was higher in 40HIGH compared with CON and 40LOW. Agitation did not impact motility (p = .581), progressive motility (p = .870), viability (p = .509) or morphology (p = .970), while long-term extender maintained higher motility (p = .002), progressive motility (p = .036), viability (p < .001) and normal acrosome (p < .001) than a short-term extender. VAP (p = .039) of 40HIGH was lower than CON in a thermo-resistance test. Neither treatment (p > .798, .766) nor agitation (p > .396, .476) impacted bacterial growth in this study. In conclusion, air contact negatively impacts boar semen pH and consequently sperm motility. Semen doses prepared with 80 or 40 ml volumes of extended boar semen with minimal air contact in the tubes yield more desirable semen quality and agitating boar semen doses daily does not have negative or positive effects on boar semen quality.

HPV infection and bacterial microbiota in the semen from healthy men.

BACKGROUND: Aberrant microbiota composition has been linked to disease development at numerous anatomical sites. Microbiota changes in reaction to viral infections, such as human papillomavirus (HPV), have been investigated almost exclusively in the female reproductive tract. However, HPV infection may also affect male health by reducing semen quality and fertility. The aim of this study was to investigate whether present HPV DNA is associated with detectable changes in semen bacterial microbiota composition and diversity. METHODS: This study relied on stored semen samples from 31 fertile healthy men who participated in the Finnish family HPV Study during the years 1998-2001. DNA was extracted from semen with PCR template preparation kit. HPV was genotyped using Luminex-based Multimetrix® assay. Microbiota was analyzed from the V3-V4 region of 16S rDNA gene following sequencing on an Illumina MiSeq platform. All statistical analyses were performed with Calypso software version 8.84. RESULTS: HPV DNA was detected in 19.4% (6/31) of the semen samples. HPV status in the semen did not impact the α-diversity estimations, as measured by Chao1 and Shannon indices, nor ß-diversity. Nevertheless, HPV-positive semen samples exhibited differences in the taxonomic composition of the bacterial microbiota including higher abundances of Moraxellaceae (p = 0.028), Streptococcus (p = 0.0058) and Peptostreptococcus (p = 0.012) compared to HPV-negative semen samples. CONCLUSION: HPV infection is associated with altered bacterial microbiota composition in semen, and this might have in impact to male health in general. As of present, it is unclear whether these changes result from HPV infection or whether altered bacterial microbiota increases susceptibility to HPV infection. More research is needed on viral-bacterial interactions in the male reproductive system.

Microbiome affecting reproductive outcome in ARTs.

Current scientific evidence reveals the importance of the human microbiome in health and disease. The presence of microbiota within the male and female reproductive tract has been well-documented and present theories imply that a possible disruption of their concentrations may have adverse effects on reproductive health and reproductive outcomes. Altered endometrial and vaginal microbiome could potential affect the reproductive outcome in infertile couples undergoing assisted reproductive techniques. Analysis of seminal fluids could also facilitate a prompt and appropriate approach in cases of abnormal male reproductive microflora. Essential knowledge on this subject could provide fertility experts better understanding with regards to unexplained fertility, increasing the success rates of ARTs. In this review, we summarise the current knowledge on the microbiota of the male and female reproductive tract and its impact on the success rates of ARTs in infertile couples.

Haemophilus seminalis sp. nov., isolated from human semen.

Two Haemophilus-like isolates with similar biochemical characteristics, designated strains SZY H1T and SZY H2, were isolated from human semen specimens. Cells were Gram-negative, non-motile, non-acid-fast, pleomorphic rods or coccobacilli. The major fatty acids (>10 %) were C16 : 0, C14 : 0, iso-C16 : 0 and/or C14 : 0 3-OH and C16 : 1 ω6c and/or C16 : 1 ω7c. The polar lipids were determined to be phosphatidylethanolamine, phosphatidylglycerol, an unidentified phospholipid, an unidentified aminophospholipid, two unidentified polar lipids and four unidentified aminolipids. The major polyamine was found to be cadaverine. The near-full-length (1462 nt) 16S rRNA gene sequences analysis showed the two isolates were nearly identical (>99.8 %), and closely matched Haemophilus haemolyticus ATCC 33390T with 98.9-99.1 % sequence similarities. Phylogenetic analysis based on 16S rRNA gene sequences and concatenation of 30 protein markers also revealed that the isolates clustered together with H. haemolyticus ATCC 33390T, and formed a distinct lineage well separated from the other members of the genus Haemophilus. Further, the average nucleotide identity values between the two isolates and their related species were below the established cut-off values for species delineation (95 %). Based on these findings, the two isolates are considered to represent a new species of the genus Haemophilus, for which name Haemophilus seminalis sp. nov. is proposed. The type strain is SZY H1T (=NBRC 113782T=CGMCC 1.17137T).

Diluente Tris suplementado com óleo de Mauritia flexuoxa sobre a qualidade do sêmen caprino após a criopreservação / Tris diluent supplemented with Mauritia flexuoxa oil on the quality of goat semen after cryopreservation

Objetivou-se verificar os efeitos, nos parâmetros espermáticos, na integridade mitocondrial, acrossomal e de membrana em células espermáticas, desencadeados pelo uso do Tris (Tris hidroximetil aminometano) suplementado com óleo de Mauritia flexuoxacomo diluente para criopreservação de sêmen caprino. Quatro caprinos clinicamente saudáveis foram utilizados. Os animais eram alimentados diariamente com volumoso (Pennisetum purpureum, Schum.), concentrado (ração peletizada com teor de 20% proteína, 300 g/animal/dia) e sal mineral específico para Caprinos (Caprinofós®), à vontade. Dois ensaios foram realizados: I ­ Teste de toxicidade; II ­ Criopreservação do sêmen com concentrações ideais. No teste de toxicidade as concentrações avaliadas foram: 5%, 10%, 15% e 20% de diluente a base de óleo de Mauritia flexuoxa. Após o teste de toxicidade, foi escolhido a concentraçãoque apresentou o melhor resultado (5%). Logo após, foram realizadas mais 32 coletas, que foram diluídas em Tris-gema-glicerol (grupo controle) ou diluente contendo óleo vegetal (Mauritia flexuoxa). As amostras foram criopreservadas com auxílio do aparelho Tk3000®. Após o período mínimo de uma semana as palhetas foram descongeladas em banho-maria a 37 °C por 30 segundos, acondicionadas em microtubos de centrifugação e homogeneizadas para a análise imediata de motilidade, vigor espermático e morfologia. Em seguida, por meio de sondas fluorescentes foram avaliadas a integridade de acrossomo, membrana plasmática (Diacetato de Carboxifluresceína e Iodeto de Propídeo) e função mitocondrial sob microscopia de epifluorescência. Quanto a motilidade e vigor, integridade mitocondrial e acrossomal, o grupo buriti foi inferior ao grupo controle. O Tris suplementado com óleo de Mauritia flexuoxa na concentração de 5% não influenciou significativamente a qualidade espermática, porém, observou-se morfologia e integridade de membrana favoráveis. Dessa forma, sendo uma alternativa para substituição de diluentes a base de produtos de origem animal.

The objective was to verify the effects, sperm parameters, mitochondrial, acrosomal and membrane integrity in sperm cells, triggered by the use of Tris (Tris hydroxymethyl aminomethane) supplemented with Mauritia flexuoxa oil as a diluent for cryopreservation of goat semen. Four goats clinically healthy were used. The animals were fed daily with bulky (Pennisetum purpureum, Schum.), concentrate (pelleted feed with 20% protein content, 300 g / animal / day) and mineral salt Specific for Goats (Caprinofós®), ad libitum. Two tests were carried out: I - Toxicity test; II - Semen cryopreservation with ideal concentrations. In the toxicity test as selected were: 5%, 10%, 15% and 20% of Mauritia flexuoxa oil-based diluent. After the toxicity test, the concentration that showed the best result (5%) was chosen. Soon after, a further 32 samples were obtained, which were diluted in Tris-glycerol (control group) or diluent containing vegetable oil (Mauritia flexuoxa). The samples were cryopreserved using the Tk3000® machine. After a minimum of one week, the samples were thawed in a 37 ° C water bath for 30 seconds, packed in centrifugation microtubes and homogenized for immediate analysis of motility, sperm vigor and morphology. Then, by means of fluorescent probes, the integrity of the acrosome, plasma membrane (Carboxyflurescein diacetate and Propidium Iodide) and mitochondrial function under epifluorescence microscopy were evaluated. As for motility and vigor, mitochondrial and acrosomal integrity, the buriti group was inferior to the control group. Tris supplemented with Mauritia flexuoxa oil at a concentration of 5% did not significantly influence sperm quality, however, favorable motility, morphology and membrane integrity were observed. Thus, being an alternative to replace diluents based on products of animal origin.

In vitro antibacterial and cytogenotoxicological properties of the aqueous extract of Cymbopogoncitratus Stapf (DC) leaf.

BACKGROUND: Microbial infection of the genital tract or semen is one of the leading causes of male infertility. Consequently, there is a need to seek alternative products from natural sources. OBJECTIVES: The antibacterial, phytochemical and cytogenotoxicological assessments of the aqueous extract of Cymbopogon citratus leaf were evaluated. METHODS: The antibacterial potential of the extract was done via agar-well diffusion and microdilution techniques. The phytochemical analysis was done via standard protocols.The cytogenotoxicity of the extract were analyzed using the Allium cepa assay. RESULTS: All test organisms were found to be sensitive to the extract except Pseudomonas. aeruginosa where no measurable zone of inhibition could be ascertained at all concentrations assessed.The highest mean inhibition diameter of 21.33±1.20mm against S. sapophyticus was recorded and a concentration-dependent susceptibility noticed. The phytochemical results revealed the presence of saponins, flavonoid, glycoside, steroids, terpenoid and alkaloids. The Alliumcepa root showed reduced mitotic indices following aconcentration-dependent increase in the extract.It can be said that the aqueous extract of C. citratus had inhibitory activities against the tested pathogenic organisms with relative anti-tumour potential. CONCLUSION: This study indicated, C. citratus could be a potential source for antibacterial compounds for the possible treatment of male reproductive related infections.

Implications of prostate inflammation on male fertility.

The prostate is the seat of three major causes of morbidity: benign prostatic hyperplasia, prostate cancer and prostatitis, three conditions in which inflammation has been implicated. A state of inflammation of the prostate gland, originally incited by an infection, an autoimmune response, a neurogenic stimulus or another trigger may have consequences on prostate functionality. In fact, male fertility depends intrinsically on the content of prostatic fluid factors secreted by the prostatic epithelium. Taking into account that the prostate gland is the major male accessory gland that exerts essential functions for male fertility, a state of local inflammation can alter male fertility by either directly impairing sperm quality or, indirectly, by causing prostate dysfunction. In the present review, we summarise the current knowledge regarding prostatitis due to well-known infections such as Escherichia coli, Chlamydia trachomatis and other commonly identified microorganisms focusing on inflammatory markers detected during these infections and seminal quality and male fertility alterations reported. We also focused on type III prostatitis or chronic nonbacterial prostatitis/chronic pelvic pain syndrome, of unknown aetiology, in which inflammation of an autoimmune origin, neurogenic stimuli or another trigger have been proposed and fertility alterations reported.

Presencia de Ochrobactrum anthropi en muestras de semen / Presence of Ochrobactrum Anthropi in Semen Samples

Introducción: El semen es una mezcla compleja de fluidos y células que posee las condiciones adecuadas para albergar microorganismos, especialmente bacterias. Objetivo: Evaluar la presencia de bacterias en el semen de individuos normozoospérmicos asintomáticos para infecciones urogenitales. Métodos: Se realizó una secuenciación estándar posterior a la amplificación por PCR con el uso de los cebadores universales 27F y 1492R para identificación de bacterias, en 10 muestras de semen de voluntarios normozoospérmicos asintomáticos para infecciones urogenitales. Resultados: Se identificó a Ochrobactrum anthropi en 8 de las 10 muestras seminales evaluadas y a Haemophilus paraurethrae o Escherichia coli en los dos restantes. O. anthropi es una bacteria comensal, ampliamente distribuida en la naturaleza, especialmente en las fuentes de agua que, a pesar de su baja virulencia, ocasionalmente causa infecciones en individuos inmunocomprometidos. Conclusión: La alta frecuencia de O. anthropi en las muestras de semen de individuos normozoospérmicos asintomáticos para infecciones urogenitales puede asociarse a procesos de contaminación durante la recolección de la muestra, debido a la amplia distribución de esta bacteria, especialmente en las fuentes de agua(AU)

Introduction: Semen is a complex combination of fluids and cells that can harbor microorganisms, especially bacteria. Objective: To assess the presence of bacteria in semen samples from asymptomatic normozoospermic individuals, for urogenital infections. Methods: Standard sequencing after PCR amplification was performed with the use of the universal primers 27F and 1492R for bacterial identification, in 10 semen samples of asymptomatic normozoospermic volunteers for urogenital infections. Results: Thisidentified Ochrobactrum anthropi in 8 out of 10 samples assessed. In the remaining two samples, we identified Haemophilus paraurethrae and Escherichia coli. O. anthropi is a commensal bacterium, widely spread in nature, especially in water sources that, despite its low virulence, occasionally cause infections in immune compromised individuals. Conclusion: The high frequency of O. anthropi in semen samples from asymptomatic normozoospermic individuals, for urogenital infections can be associated with contamination during the collection of the sample, due to the wide distribution of this bacterium, especially in water sources(AU)

Review: Risks of disease transmission through semen in cattle.

The purpose of this paper is to review scientific evidence concerning pathogens that could potentially be transmitted via bovine semen. As a result of a careful analysis of the characteristics of infections that may cause transmission of disease through semen, effective control procedures can be identified that provide minimal constraint to the introduction of new bulls into herds for natural breeding and importation of valuable novel genetics through artificial insemination. The potential for transmission through bovine semen and corresponding effective control procedures are described for bovine herpesvirus 1, bovine viral diarrhea virus, bovine leukemia virus, lumpy skin disease virus, bluetongue virus, foot-and-mouth disease virus, and Schmallenberg virus. Brief consideration is also provided regarding the potential for transmission via semen of Tritrichomonas foetus, Campylobacter fetus venerealis, Brucella abortus, Leptospira spp., Histophilus somni, Ureaplasma diversum, Mycobacterium avium subsp. paratuberculosis, Chlamydiaceae, Mycobacterium bovis, Coxiella burnetii, Mycoplasma mycoides ssp. mycoides and Neospora caninum. Thoughtful and systematic control procedures can ensure the safety of introducing new bulls and cryopreserved semen into cattle production systems.

Molecular detection of Chlamydia trachomatis and semen quality of sexual partners of infertile women.

Chlamydia trachomatis is considered as the bacterium that is more sexually transmitted as cause of male urethritis, epididymitis, orchitis and infertility. A total of 116 semen samples of men whose couples are infertile women were analysed. The quality of the semen was measured by standard procedures recommended by WHO while C. trachomatis was detected by the PCR assay. Thirty-seven semen samples were positive for C. trachomatis (31.9%). Regarding semen analysis, no different values were observed between positive and negative samples to C. trachomatis. However, the presence of leucocytes and erythrocytes suggests an inflammatory process; however, these were high in negative samples to C. trachomatis. Furthermore, an association between low seminal volume at 1, 5 ml and the positivity to C. trachomatis was observed (OR=2, 1; CI95 % 1,16-3,07). The total semen volume is a contribution by the various accessory glands (this reflects the secretory activity of the glands); a low semen volume could be due to an obstruction of the ejaculatory duct or infection of accessory glands by C. trachomatis. More studies are necessary to identify the causes of a reduced semen volume.

Molecular characterization of ureaplasmas isolated from reproductive tract of goats and sheep from Brazil

Abstract Ovine/caprine ureaplasmas have not yet been assigned a species designation, but they have been classified into nine serotypes. Herein ureaplasmas were searched for in 120 samples of vulvo vaginal mucous from sheep and 98 samples from goats at 17 farms. In addition, semen samples were collected from 11 sheep and 23 goats. The recovered ureaplasma were from sheep and goats from animals without any reproductive disorder symptoms, but not all animals presented positive cultures. In sheep, 17 (68%) cultures of vulvovaginal mucous were positive for ureaplasma and 11 (27%) samples of semen presented positive cultures in animals with clinical signs of orchitis, balanoposthitis or low sperm motility. In goats four ureaplasma isolates were obtained from vulvovaginal mucus, but the semen samples were all negative. The isolates were submitted to Pulsed-field gel electrophoresis methodology and their 16S rRNA genes were sequenced. Fifty percent of ureaplasma recovered from sheep allowed for PFGE typing. Eleven isolates showed eight profiles genetically close to the bovine ureaplasmas. The 16S rRNA gene sequencing showed differences or similarities of isolates from sheep and goats, and the reference strains of bovine and human ureaplasma. Four clinical isolates from sheep were grouped separately. The studied ureaplasma isolates showed to be a diverse group of mollicutes.

Do antimicrobial peptides PR-39, PMAP-36 and PMAP-37 have any effect on bacterial growth and quality of liquid-stored boar semen?

The use of antimicrobial peptides (AMP) has become one of the most promising alternatives to the use of antibiotics (Abs) in semen extender's formulation to overcome the increasing bacterial resistance to antibiotics. However, AMP may impair boar sperm quality, so that their deleterious effects might be higher than their effectiveness against bacteria. Thus, the aim of this study was to determine whether three different AMP, the proline-arginine-rich antimicrobial peptide PR-39 (PR-39), and the porcine myeloid antimicrobial peptides 36 (PMAP-36) and 37 (PMAP-37) had any effect upon boar sperm quality and bacterial growth. For this purpose, three different concentrations of each peptide (1 µM, 10 µM and 20 µM for PR-39 and 0.5 µM, 1 µM and 3 µM for PMAP-36 and PMAP-37) were added to 2 mL of a pool of extended semen with BTS without Abs; two controls, one without AMPs and Abs, and the other with Abs only were used for each peptide (n = 3). Total (TMOT) and progressive (PMOT) sperm motility, sperm viability and bacterial concentration were assessed before the addition of each AMP or Abs and at 1, 3, 6, 8 and 10 days post-addition. For each AMP, results revealed a drop in the TMOT and PMOT in all treatments and controls. In regard to sperm viability, while PR-39 at 10 µM maintained it in values similar to those of the control with Abs and PMAP-36 kept also the sperm viability in a similar fashion to the treatment with Abs, PMAP-37 was more effective in keeping sperm viability than controls (P < 0.05). Whereas PR-39 at 20 µM and PMAP-37 at 3 µM were quite effective in controlling bacterial load, PMAP-36 did not avoid bacterial growth at any concentration tested. In conclusion, taking all results together, PMAP-37 seems to be a suitable candidate to replace Abs in extended semen, as it hardly impairs sperm viability and controls the bacterial load. Nevertheless, further studies are still required to improve its effectiveness.

Discovery of a Novel Seminal Fluid Microbiome and Influence of Estrogen Receptor Alpha Genetic Status.

Bacteria harbored in the male reproductive system may influence reproductive function and health of the male and result in developmental origins of adult health and disease (DOHaD) effects in his offspring. Such effects could be due to the seminal fluid, which is slightly basic and enriched with carbohydrates; thereby, creating an ideal habitat for microbes or a potential seminal fluid microbiome (SFM). Using wild-type (WT) and estrogen receptor-alpha (ESR1) knockout (KO) male mice, we describe a unique SFM whose inhabitants differ from gut microbes. The bacterial composition of the SFM is influenced according to whether mice have functional Esr1 genes. Propionibacterium acnes, causative agent of chronic prostatitis possibly culminating in prostate cancer, is reduced in SFM of ESR1 KO compared to WT mice (P ≤ 0.0007). In certain genetic backgrounds, WT mice show a greater incidence of prostate cancer than ESR1 KO, which may be due to increased abundance of P. acnes. Additionally, select gut microbiome residents in ESR1 KO males, such as Lachnospiraceae and Christensenellaceae, might contribute to previously identified phenotypes, especially obesity, in these mutant mice. Understanding how genetics and environmental factors influence the SFM may provide the next frontier in male reproductive disorders and possibly paternal-based DOHaD diseases.

Real-time PCR for detection of Brucella ovis and Histophilus somni in ovine urine and semen / PCR em tempo real para detecção de Brucella ovis e Histophilus somni em urina e sêmen ovino

A epididimite infecciosa ovina é uma das principais enfermidades reprodutivas de carneiros. O presente estudo teve por objetivo desenvolver protocolos de PCR em tempo real para B. ovis e H. somni e avaliar sua aplicabilidade em amostras de sêmen e urina de carneiros. Delinearam-se primers e sondas espécie-específicos para cada agente. As sondas foram delineadas com o sistema TaqMan incorporando um marcador FAM para B. ovis e Cy5 para H. somni na extremidade 5' e um quencher na extremidade 3'. A PCR em tempo real para B. ovis e H. somni foi altamente sensível, uma vez que a amplificação de DNA ocorreu com até 0,2ng de DNA/reação. A especificidade dos iniciadores e sondas foi avaliada com amostras de DNA de outros agentes causadores de epididimite ovina e nenhuma amplificação inespecífica foi observada. A aplicabilidade da técnica em amostras biológicas também foi confirmada, pois não houve perda de eficácia (P>0,05) quando comparada à PCR convencional com amostras de sêmen e urina de carneiros experimentalmente infectados.

Efecto de los factores solubles de Staphylococcus aureus, Staphylococcus capitis y Staphylococcus epidermidis sobre la funcionalidad espermática / Effect of soluble factors of Staphylococcus aureus, Staphylococcus capitis and Staphylococcus epidermidis on sperm functionality

ANTECEDENTES: La interacción entre los espermatozoides con algunas especies bacterianas o sus factores solubles influyen en el deterioro de la calidad seminal, alterando la función reproductiva del hombre. OBJETIVO: El objetivo de este trabajo fue determinar el efecto de los factores solubles de Staphylococcus aureus, Staphylococcus capitis y Staphylococcus epidermidis sobre la calidad seminal. MÉTODO: Los factores solubles producto del metabolismo bacteriano de las cepas de S. aureus y S. Capitis sensible a oxacilina y S. aureus y S. Epidermidis resistente a oxacilina se incubaron con las muestras de semen de 20 voluntarios y se cuantificaron los parámetros seminales convencionales y funcionales por microscopía y citometría de flujo, respectivamente. RESULTADOS: Se observó una disminución en la movilidad espermática con los factores solubles de S. aureus, esta disminución fue mayor con la cepa sensible y el efecto negativo sobre la movilidad fue inmediato. Al incubar los espermatozoides con los factores solubles de S. aureus sensible a oxacilina, se afectaron todos los parámetros funcionales excepto la integridad de la cromatina y se observó menor liberación de especies reactivas de oxígeno; con los factores solubles de la cepa de S. aureus resistente a oxacilina se observó una disminución en la lipoperoxidación de membrana y en la expresión de anexina V. CONCLUSIÓN: Este estudio da cuenta del efecto negativo de los factores solubles de la bacteria S. aureus tanto sensible como resistente a oxacilina sobre los parámetros espermáticos convencionales y funcionales, y por ende en su función reproductiva.

BACKGROUND: The interaction between sperm with some bacteria species and their soluble factors are the deterioration of semen quality by altering the reproductive function of man. AIM: The aim of this study was to determine the effect of soluble factors Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus capitis on semen quality. METHODS: The soluble factors product of bacterial metabolism of the strains of S. aureus and S. capitis methicillin sensitive and S. aureus and S. epidermidis resistant to oxacillin, were incubated with semen samples from 20 volunteers. Subsequently, conventional seminal parameters were measured and functional quantified by microscopy and flow cytometry, respectively. RESULTS: A decrease was observed in sperm motility with soluble factors of S. aureus, this decrease was higher with the sensitive strain that with oxacillin resistant strain and the negative effect on motility was immediate. By incubating the sperm with soluble factor from oxacillin-sensitive S. aureus, all functional parameters were affected except the chromatin integrity and reduced release of reactive oxygen species, mean fluorescence intensity in oxacillin resistant S. aureus strain was decrease in membrane lipid peroxidation and annexin V expression. CONCLUSIONS: This study reports the negative effect of soluble factors of bacteria either S. aureus sensitive and resistant to oxacillin, over conventional and functional sperm parameters, and therefore in their reproductive function.

Challenges and Limits Using Antimicrobial Peptides in Boar Semen Preservation.

Antibiotics are of great importance for the preservation of ejaculates for livestock breading. The use of antibiotics, however, is not an appropriate compensation for a lack of hygiene standards in artificial insemination (AI) centres. Sophisticated hygiene management and the proper identification of hygienic critical control points (HCCPs) at AI centres provide the basis for counteracting the development of antibiotic resistance in contaminant bacteria and their settlement in AI centres. In recent years, efforts have been made to use antimicrobial peptides (AMPs) in the preservation of boar semen. Investigations have included the testing of synthetic magainin derivatives and cyclic hexapeptides. One prerequisite for the application of AMPs is that they have a minor impact on eukaryotic cells. Bacterial selectivity, proteolytic stability, thermodynamic resistance, and mechanisms including synergistic interaction with conventional antibiotics have made cyclic hexapeptides highly promising candidates for potential application as peptide antibiotics for semen preservation.