Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production.

Sensitivity to freezing remains a critical issue in stallion semen cryopreservation procedures. To explore this topic in-depth, semen was collected from ten stallions, diluted with three different extenders, transported to the laboratory, and then centrifuged and frozen with four different extenders. We conducted analyses of sperm kinetics, mitochondrial membrane potential (MMP), and hydrogen peroxide content both before and after freezing. Additionally, we assessed antioxidant activity using the ABTS and FRAP methods and measured nitric oxide stable metabolites (NOx) in the blank extenders, seminal plasma, and extenders conditioned by spermatozoa before and after freezing. We found significant variability in the antioxidant activity and NOx content of the blank extenders and the seminal plasma. In the seminal plasma, ABTS-based antioxidant activity and NOx values were correlated with some sperm kinematic parameters and MMP in refrigerated semen, while no correlation was observed in frozen sperm parameters. Sperm function varied significantly between stallions but not between extenders, either before or after freezing. However, significant differences in antioxidant activities and NOx values were found among extenders conditioned following freezing. These results provide new insights into the factors contributing to the variability in individual stallions' tolerance to sperm freezing.

Polymer chain extenders induce significant toxicity through DAF-16 and SKN-1 pathways in Caenorhabditis elegans: A comparative analysis.

Polymer chain extenders, commonly used in plastic production, have garnered increasing attention due to their potential environmental impacts. However, a comprehensive understanding of their ecological risks remains largely unknown. In this study, we employed the model organism Caenorhabditis elegans to investigate toxicological profiles of ten commonly-used chain extenders. Exposure to environmentally relevant concentrations of these chain extenders (ranging from 0.1 µg L-1 to 10 mg L-1) caused significant variations in toxicity. Lethality assays demonstrated the LC50 values ranged from 92.42 µg L-1 to 1553.65 mg L-1, indicating marked differences in acute toxicity. Sublethal exposures could inhibit nematodes' growth, shorten lifespan, and induce locomotor deficits, neuronal damage, and reproductive toxicity. Molecular analyses further elucidated the involvement of the DAF-16 and SKN-1 signaling pathways, as evidenced by upregulated expression of genes including ctl-1,2,3, sod-3, gcs-1, and gst-4. It implicates these pathways in mediating oxidative stress and toxicities induced by chain extenders. Particularly, hexamethylene diisocyanate and diallyl maleate exhibited markedly high toxicity among the chain extenders, as revealed through a comparative analysis of multiple endpoints. These findings demonstrate the potential ecotoxicological risks of polymer chain extenders, and suggest the need for more rigorous environmental safety assessments.

Effects of different extenders, storage temperatures, and antioxidant supplementation on chilled semen quality: a review.

The successful preservation of ram semen is essential to promote genetic variability, ensure semen transportation, and inseminate multiple ewes. Currently, either animal or plant-based lipoprotein-based extenders are used for semen preservation. Animal product-based extenders include milk and egg yolk, while soybean lecithin is a plant-based extender. Although extenders containing products of animal origin better preserve the quality of chilled semen, the in vivo efficacy after 24 h of storage is still of great concern. Storage temperature is another important and effective factor in preserving sperm quality, whereby different storage temperatures are adopted to enhance the storage life of sperm. Low temperatures (4-5 °C) better preserve sperm quality for a longer duration than high temperatures (15, 20, and 25 °C). Moreover, antioxidant supplementation has a positive impact on sperm quality during liquid storage. The current review summarizes the outcomes of various extenders, different storage temperatures, and antioxidant supplementation on the liquid storage of ram sperm.

Evaluating attributes of a collaborative model of service delivery for hereditary cancer risk assessment.

The purpose of this nonrandomized study was to compare several attributes of hereditary cancer risk assessment using a collaborative model of service delivery. Arm 1 included patients seen in-person by a board-certified genetic counselor (CGC), Arm 2 included high-complexity triaged patients from distant sites who received telegenetics with a CGC, and Arm 3 included low-complexity triaged patients from distant sites who had in-person risk assessment with a locally placed genetic counselor extender (GCE). A total of 152 patients consented and 98 had complete data available for analysis (35 in Arm 1, 33 in Arm 2, and 30 in Arm 3). The three groups were comparable in age, ethnicity, education, employment, and cancer status. There was no significant difference in median wait time or distance traveled to receive care across all three arms. However, if patients in Arms 2 and 3 had to access the CGC in-person, they would have had to travel significantly further (p < 0.0001). The time spent in a session was significantly longer in Arm 3 with a GCE than with a CGC in-person or by telegenetics (p < 0.01). There was no difference in the number of essential elements covered in the appointment, change in cancer worry, or appointment satisfaction across all three arms, although the sample size was small. Employing a collaborative model of service delivery with GCEs and telegenetics is feasible, satisfactory to patients and reduces the distance patients travel to access hereditary cancer genetic services.

Bull Semen Obtained on Beef Farms by Electroejaculation: Sperm Quality in the First Two Hours of Storing with Different Extenders and Holding Temperatures.

Sperm quality decreases over time, so bull semen may need to be preserved after field collection. However, the effect of handling such semen samples from commercial farms and placing them in very short-term storage has not been elucidated. Therefore, ejaculate from 25 bulls from 1 dairy and 14 beef cattle farms were collected under farm conditions and evaluated for semen quality during the first two hours after collection. Two commercial extenders (AndroMed® and BIOXcell®) and two different storage temperatures (5 °C and room temperature) were used to evaluate the influence on semen quality and sperm kinetics in ejaculates grouped into three evaluation times, based on time since collection (Time 1: <75 min, n = 7; Time 2: 75-105 min, n = 11; and Time 3: 105-120 min, n = 7). Classical semen parameters, sperm motion kinetics by CASA and colony-forming units were assessed. The differences between both extenders in curvilinear and straight-line velocities (VCL and VSL) for the different time groups (Time 2 and Time 3) were statistically significant for p < 0.05. AndroMed® showed lower VSL, straightness and linearity in sperm compared to BIOXcell® (p < 0.05). In conclusion, AndroMed® induced more curvilinear movement, while BIOXcell® stimulated straighter motility.

Effects of extender type and storage time on sperm quality parameters of Kail ram semen stored at 5 °C.

The study was designed to evaluate the effect of different extenders and storage times on sperm quality parameters of extended Kail ram semen. Semen was collected from five adult Kail rams using an artificial vagina. Semen samples with >70% total sperm motility were pooled, diluted with Tris (TR), sodium citrate (SC), and skim milk (SM)-based extenders, and stored at 5 °C. Sperm motility and kinematics, viability, and plasma and acrosomal membrane integrity were assessed every 24 hrs for 120 hrs. Sperm longevity was estimated by placing semen in a water bath at 37°C to assess sperm kinematics. Storage time as the main effect had a negative (P < 0.05) impact on sperm quality parameters. The percentages of total motile (TM), progressive motile (PM), rapid, and medium progressive (MP) motile sperm were similar at 24 hrs of storage for the three extenders. However, semen extended in TR had a higher (P < 0.05) percentage of TM, PM, rapid, and MP sperm compared to SM and SC extenders at 48 hrs of storage. The sperm kinematics (VCL, VSL, VAP, and ALH) of progressive sperm were similar for all three extenders up to 72 hrs of storage. In addition, semen extended in TR had a higher (P < 0.05) percentage of sperm with intact plasma and acrosomal membranes at a storage time of 48 hrs. At 37 °C, the percentage of TM and PM sperm was higher (P < 0.05) in the TR extender than in the SM and SC extenders at 60 minutes and beyond. In conclusion, the Kail ram sperm extended in TR and stored at 5 °C maintained better quality after 48 hrs storage than sperm extended in SM and SC extenders. At 37 °C, sperm extended in TR also retained better quality parameters at 60 min and afterward than sperm extended in SM and SC.

Antimicrobial Resistance in Vaginal Bacteria in Inseminated Mares.

Antimicrobials are added to semen extenders to inhibit the growth of bacteria that are transferred to the semen during collection. However, this non-therapeutic use of antimicrobials could contribute to the development of antimicrobial resistance. The objective of this study was to determine changes in the antibiotic susceptibility of vaginal microbiota after artificial insemination. Swabs were taken from the vagina of 26 mares immediately before artificial insemination and again 3 days later. Bacteria isolated from the vagina at both time points were subjected to antibiotic susceptibility testing and whole-genome sequencing. In total, 32 bacterial species were identified. There were increases in the resistance of Escherichia coli to trimethoprim (p = 0.0006), chloramphenicol and (p = 0.012) tetracycline (p = 0.03) between day 0 and day 3. However, there was no significant effect of exposure to antibiotics in semen extenders with respect to the resistance of Staphylococcus simulans and Streptococcus equisimilis (p > 0.05). Whole-genome sequencing indicated that most phenotypic resistance was associated with genes for resistance. These results indicate that the resistance patterns of vaginal bacteria may be affected by exposure to antibiotics; therefore, it would be prudent to minimize, or preferably, avoid using antibiotics in semen extenders.

Inflatable penile prosthesis placement after prior transcorporal artificial urinary sphincter placement: A case report.

Stress urinary incontinence and erectile dysfunction often coexist in men surgically treated for prostate cancer. Despite many men having both an artificial urinary sphincter and inflatable penile prosthesis to treat these conditions, there is limited information in the literature to guide surgeons when it comes to placing both devices. We recommend obtaining direct exposure of proximal crura to allow for complete dilation of corporal spaces for proper prosthetic placement. Further dissection via penoscrotal incision or perineal counter-incision can be utilized. Surgeons should consider dorsal lithotomy position at time of IPP placement to allow for perineal exposure.

Effect of Aromatic Chain Extenders on Polyurea and Polyurethane Coatings Designed for Defense Applications.

The present work describes the synthesis of new versatile polyurea (PU) and polyurethane (PUR) matrices, including different chain extenders, which facilitate the design of distinct, tunable properties, and high-performance derivatives. These polymers can be used for various defense and security applications, such as coatings for ballistic protection, CBRN protection, binders for energetic formulations, etc. Combining aliphatic and aromatic molecules in PU or PUR structures enables the synthesis of polymers with improved and controllable thermo-mechanical properties. Thus, for polyurea synthesis, we utilized two types of polymeric aliphatic diamines and three types of aromatic chain extenders (1,1'-biphenyl-4,4'-diamine, benzene-1,2-diamine, and 1,2-diphenylhydrazine). An analogous method was used to synthesize polyurethane films by employing one polymeric aliphatic polyol and three types of aromatic chain extenders (benzene-1,3-diol, benzene-1,4-diol, and benzene-1,2,3-triol). Subsequently, various analytic techniques (Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR), single cantilever dynamic mechanical analysis (DMA), thermogravimetric analysis (TGA), frequency-dependent shear modulus survey, tensile tests, water contact angle measurements, and scanning electron microscopy (SEM) with energy-dispersive X-ray analysis (EDX)) have been utilized to characterize the synthesized materials and to evaluate the influence of each chain extender on their final properties.

Effect of freeze-dried quail egg white and yolk addition to semen extender on viability of rooster sperm stored for 6 h at 4°C.

The effect of freeze-dried quail egg white and yolk addition to basic EK extender on morphology and motility of chicken broiler breeder semen was investigated. Fresh pooled semen was divided into eight parts: fresh, undiluted (control), diluted in 1:2 ratio (v/v) with basic EK extender, EK + 200 mg/ml of egg white, EK + 100 mg/ml of egg white, EK + 50 mg/ml of egg white, EK + 100 mg/ml of egg yolk, EK + 50 mg/ml of egg yolk, EK + 25 mg/ml of egg yolk. Semen samples were evaluated 15 min after dilution and after 6 h of storage at 4°C. In the fresh semen, the number of live normal sperm was the highest in semen diluted with EK + 200 mg of egg white and EK + 100 mg of egg yolk, while the highest sperm motility was in the neat semen. Semen storage reduced the number of normal sperm in all analysed semen samples. In the neat semen, the number of normal sperm decreased, in relation to the fresh not-stored samples, by 36.8% (from 72.3% to 35.5%), with EK extender by 9.2%, in samples enriched with egg white, from 8.4% (EK + 200 mg) to 10.0% (EK + 100 mg), and in EK with egg yolk addition, from 1.2% (EK + 50 mg) to 10.6% (EK + 100 mg). The highest percentage of motile sperm was observed in EK extender enriched with 50 mg of egg white (77.1%) and EK + 25 mg of egg yolk (65.3%).

Design and Assessment of a Pharmacy Student-Delivered Preceptor Development Program.

Objective. To design and assess the use of a pharmacy student-delivered preceptor development program.Methods. A student-delivered preceptor development program was developed to ensure all preceptors received documented preceptor development. A menu of discussion topics and associated teaching sheets were created by the school's office of experiential education. On each rotation, advanced pharmacy practice experience (APPE) students led discussions with their preceptors on a topic chosen by the preceptor and submitted documentation of the education. Preceptors answered a survey related to the amount of information and time required for the program, their preference for different formats of preceptor development, one important thing they learned through the program, and future development topics of interest. Students were interviewed regarding their perceptions and use of the program.Results. A novel student-delivered preceptor development program resulted in documentation of preceptor development for all APPE rotations. Seventy-nine preceptors (31% response rate) participated in the survey. In their responses, preceptors generally agreed that they were able to customize their development and incorporate what they learned into practice, and that the program had a convenient format and was valuable for preparing students to be future preceptors. Students reported that the program improved their confidence in communicating with a supervisor and prepared them for precepting.Conclusion. A student-delivered preceptor development program improved documented preceptor development. The program allowed preceptors to customize development opportunities in a convenient format. It was perceived positively by preceptors who would recommend the program to other schools of pharmacy.

Preparation and Characterization of Water-borne Polyurethane Based on Benzotriazole as Pendant Group with Different N-Alkylated Chain Extenders and Its Application in Anticorrosion.

A series of novel anti-corrosive coatings were synthesized successfully. Water-borne polyurethane (WPU) was synthesized using polyethylene glycol and modified by grafting benzotriazole (BTA) as a pendant group (WPU-g-BTA) and N-alkylated amines (ethylene diamine (A), diethylene triamine (B), triethylene tetramine (C)) as side-chain extenders. Fourier-transform infrared spectroscopy, thermogravimetry, and dynamic mechanical analyses were used to characterize the structural and thermomechanical properties of the samples. A gas permeability analyzer (GPA) was used to evaluate molecular barrier properties. The corrosion inhibition performance of WPU-g-BTA-A, WPU-g-BTA-B, and WPU-g-BTA-C coatings in 3.5 wt% NaCl solution was determined by electrochemical measurements. WPU-g-BTA-C coating synthesized with a high cross-linking density showed superior anticorrosive performance. The as-prepared coatings exhibited a very low icorr value of 0.02 µA.cm-2, a high Ecorr value of -0.02 V, as well as excellent inhibition efficiency (99.972%) and impedance (6.33 Ω) after 30 min of exposure.

Use of commercial extenders, with and without the addition of egg yolk, for cooling llama semen.

The objective of this study was to evaluate the effect of two commercial extenders, AndroMed® (AM) and Androstar® Plus (AS) both with and without the addition of egg-yolk (EY), for cooling llama semen. A total of sixteen ejaculates were collected from four males. Each ejaculate was divided into four aliquots and diluted with: AM, AM with 20 % EY (AM-EY), AS and AS with 20 % EY (AS-EY) and then cooled to 5 °C in an Equitainer®. Evaluations were carried out in raw semen, after dilution (0 h) and after 24 and 48 h of cooling. Data were analysed using either Friedman or ANOVA. Although total motility decreased in all cooled samples compared to the corresponding 0 h (P < 0.05), the highest percentages were observed in AM-EY being significantly higher than all other cooled samples after 24 h and higher than AS and AS-EY after 48 h. No significant differences were observed in the percentages of live acrosome-intact sperm between extenders at all times tested. A significant decrease in the percentage of sperm membrane osmotic function was observed in samples cooled with AS and AS-EY after 24 and 48 h vs. raw semen and in AM 48 h vs. raw semen. Finally, a significant increase in the percentage of sperm with abnormal tails was observed in the samples cooled with AS and AS-EY. Of all the extenders used, AndroMed® could be considered an option for cooling llama semen and the addition of EY to this extender improves its effectiveness. DATA AVAILABILITY: The data that support the findings of this study are available from the corresponding author upon reasonable request.

Sperm quality and morphometry characterization of cryopreserved canine sperm in ACP-106c or TRIS.

Morphological sperm evaluation supported by the morphometry can be used in the determination of the seminal quality and in the investigation of potential extenders. Although there are studies comparing TRIS and ACP extenders, there are no comparative studies between them for the computerized assisted semen analysis (CASA), sperm viability, membrane functionality and sperm morphometry parameters of cryopreserved canine semen. Hence, we aimed to evaluate the effects of ACP-106c and TRIS on post-freezing canine sperm quality. Five dogs were submitted to semen collection twice with one-week interval. The semen was evaluated within the parameters: total motility, vigor, concentration, viability, plasma membrane functionality, morphology and morphometry. In the morphometric evaluation, the morphologically normal sperm was measured as: length, width, area and perimeter of the head and the midpiece, tail length and total length. The parameters of ellipticity, elongation, regularity and roughness were determined. Then, the semen was divided into two aliquots that were diluted in TRIS or ACP-106c, with the addition of egg yolk and glycerol. The diluted semen was refrigerated and frozen. The thawed samples were evaluated. Total motility, viability, sperm membrane functionality and normal morphology reduced after thawing in both extenders (morphology reduced from 89.60 ± 1.3% to 84.40 ± 1.8 and 84.60 ± 1.1% in TRIS and ACP-106c, respectively). However, it did not differ between TRIS and ACP-106c. In the ACP-106c the sperm head defects in cryopreserved semen were higher compared to fresh semen (P < 0.05). For all the morphometric parameters evaluated, there were no differences between fresh and cryopreserved samples (3.70 ± 0.4% vs. 2.30 ± 0.5%). In kinetics, with an interval of one week statistical differences between the extenders were found only in the parameters ALH and LIN (P < 0.05). Regardless of the extender, there were no changes in the morphometric parameters of sperm after thawing.

Self-Healable, Strong, and Tough Polyurethane Elastomer Enabled by Carbamate-Containing Chain Extenders Derived from Ethyl Carbonate.

Commercial diol chain extenders generally could only form two urethane bonds, while abundant hydrogen bonds were required to construct self-healing thermoplastic polyurethane elastomers (TPU). Herein, two diol chain extenders bis(2-hydroxyethyl) (1,3-pheny-lene-bis-(methylene)) dicarbamate (BDM) and bis(2-hydroxyethyl) (methylenebis(cyclohexane-4,1-diy-l)) dicarbamate (BDH), containing two carbamate groups were successfully synthesized through the ring-opening reaction of ethylene carbonate (EC) with 1,3-benzenedimetha-namine (MX-DA) and 4, 4'-diaminodicyclohexylmethane (HMDA). The two chain extenders were applied to successfully achieve both high strength and high self-healing ability. The BDM-1.7 and BDH-1.7 elastomers had high comprehensive self-healing efficiency (100%, 95%) after heated treatment at 60 °C, and exhibited exceptional comprehensive mechanical performances in tensile strength (20.6 ± 1.3 MPa, 37.1 ± 1.7 MPa), toughness (83.5 ± 2.0 MJ/m3, 118.8 ± 5.1 MJ/m3), puncture resistance (196.0 mJ, 626.0 mJ), and adhesion (4.6 MPa, 4.8 MPa). The peculiar mechanical and self-healing properties of TPUs originated from the coexisting short and long hard segments, strain-induced crystallization (SIC). The two elastomers with excellent properties could be applied to engineering-grade fields such as commercial sealants, adhesives, and so on.

The Natural Cryoprotectant Honey for Fertility Cryopreservation.

Honey is a mixture of 25 sugars with other bioactive substances (i.e., organic acids, enzymes, antioxidants, and vitamins) and has been known as a highly nutritious functional food. Traditionally, it has been widely used in medicinal applications to cure various diseases. The effectiveness of honey in different applications has been used for its antimicrobial activity, absorption of hydrops, cleansing, removing odor, assisting granulation, recovery of nutrition, and formation of tissue and epithelium, which proved that honey has dehydrating and preserving properties to make it ideal for the cryopreservation of cells and tissues. Cryopreservation is an advanced preservation technique for tissue, cells, organelles, or other biological specimen storage, performed by cooling the sample at a very low temperature. It is the most common approach to improved preserving fertility (sperm, embryos, and oocytes) in different species that may undergo various life-threatening illnesses and allows for the genetic screening of these cells to test the sample for diseases before use. However, with toxic cryoprotectant (CPA), cryopreservation of fertility has been challenging because of their particular structure and sensitivity to chilling. Honey's unique composition, as well as its dehydrating and preserving properties, qualify it to be used as a natural cryoprotectant. The aim of this study is to emphasize the ability of honey as a natural cryoprotectant in cryopreservation. The articles for this review were searched from Google Scholar, PubMed, Science Direct, Web of Science, and Scopus, using the keywords, honey, cryopreservation, natural cryoprotectant/CPAs, extenders, and fertility. Honey, as a natural cryoprotectant in fertility cryopreservation, yielded satisfactory results, with respect to improved post-thaw quality and viability. It is now proved as a non-toxic and highly efficient natural cryoprotectant in fertility preservation because its increasing viscosity at low temperature can provide a protective barrier to cells by reducing ice formation. Furthermore, its antioxidant property plays a vital role in protecting the cells from thermal damage by reducing the reactive oxygen species (ROS). This review provides a road map for future studies to investigate the potential of honey in the cryopreservation of other cells and tissue and contribute to the scientific research, regarding this remarkable natural product.

Analysis of clinical and radiological outcomes of long tibial stemmed total knee arthroplasty in knee osteoarthritis complicated by tibial stress fracture.

OBJECTIVE: Knee arthritis associated with tibial stress fractures represents an uncommon and difficult clinical scenario to treat. The use of long, fluted tibial extension rods has been vital in the management of such cases owing to immediate fracture stability and single-stage surgery without the need to open the fracture site. This study investigates clinical and radiological outcomes following total knee arthroplasty using a tibial extension stem in cases of knee osteoarthritis with tibial stress fracture. METHODS: From February 2015 to December 2020, 17 patients who had total knee arthroplasty implanted with a long stemmed tibial component were included in the study. Patient data were analyzed for knee range of motion, deformities, Knee Society score, knee function score, and time to fracture union in the pre- and postoperative periods. RESULTS: The mean follow-up duration was 22.7 ± 11.68 months (range 12-60 months), and mean time to fracture healing was 10.23 ± 2.81 weeks (range 8-20 weeks). The preoperative mean fixed flexion deformity improved from 8.53 ± 3.43° to a mean of 0.29°, and knee flexion improved from 79.4 ± 13.90° to 125.29 ± 8.74° on postoperative assessment. The Knee Society score improved from a mean preoperative score of 18.94 ± 5.55 (range 8-28) to 89.41 ± 7.5 (range 74-102, p value < 0.001). Similarly, the knee function score improved significantly from a mean preoperative score of 15.5 ± 4.48 (range 8-26) to a mean of 85 ± 6.09 (range 72-94, p value < 0.001). CONCLUSION: Total knee arthroplasty using long tibial extenders has been an effective and safe surgical option for patients with advanced osteoarthritis with tibial stress fractures.

The Stallion Spermatozoa: A Valuable Model to Help Understand the Interplay Between Metabolism and Redox (De)regulation in Sperm Cells.

Significance: Proper functionality of the spermatozoa depends on the tight regulation of their redox status; at the same time these cells are highly energy demanding and in the energetic metabolism, principally in the electron transport chain in the mitochondria, reactive oxygen species are continuously produced, in addition to that observed in the Krebs cycle and during the ß-oxidation of fatty acids. Recent Advances: In addition, in glycolysis, elimination of phosphate groups from glyceraldehyde 3-phosphate and dihydroxyacetone phosphate results in the byproducts glyoxal (G) and methylglyoxal (MG); these products are 2-oxoaldehydes. The presence of adjacent carbonyl groups makes them strong electrophiles that react with nucleophiles in proteins, lipids, and DNA, forming advanced glycation end products. Critical Issues: This mechanism is behind subfertility in diabetic patients; in the animal breeding industry, commercial extenders for stallion semen contain a supraphysiological concentration of glucose that promotes MG production, constituting a potential model of interest. Future Directions: Increasing our knowledge of sperm metabolism and its interactions with redox regulation may improve current sperm technologies in use, and shall provide new clues to understanding infertility in males. Moreover, stallion spermatozoa due to its accessibility, intense metabolism, and suitability for proteomics/metabolomic studies may constitute a suitable model for studying regulation of metabolism and interactions between metabolism and redox homeostasis. Antioxid. Redox Signal. 37, 521-537.

Antibiotic Resistance Patterns in Cervical Microbes of Gilts and Sows.

Extenders for boar semen contain antibiotics, which may induce antimicrobial resistance (AMR) in inseminated females. The objective was to investigate AMR of bacteria isolated from the cervix of sows and gilts in standing heat, representing females previously exposed to antibiotics in the semen extender and non-exposed females, respectively. Cervical swabs were taken from 30 multiparous sows and 30 gilts prior to their first insemination. After culturing on agar plates, bacterial isolates were identified by Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry and antimicrobial minimum inhibitory concentrations (MIC) were determined. Differences in antibiotic resistance between sows and gilts were analyzed by Chi-squared or Fisher's exact test. Bacteria isolated were mostly Staphylococcus spp., Streptococcus spp. and Corynebacterium spp. Higher MICs were observed for isolates from sows than from gilts. Most (>80%) Corynebacterium spp. were resistant to clindamycin; small numbers (<20%) were resistant to gentamicin, penicillin, vancomycin, ciprofloxacin and rifampicin, with no differences between gilts and sows. Corynebacterium from gilts were more often resistant to tetracycline than those from sows (25% vs. 4.17%; p = 0.04). In conclusion, bacteria from the porcine cervix showed low resistance to most antibiotics except for clindamycin, but antibacterial resistance may increase with increasing parity.

Semen collection, evaluation, and cryopreservation in the bonobo (Pan paniscus).

BACKGROUND: Captive breeding of bonobos (Pan paniscus) has proven to be successful, but maintaining genetic diversity remains a challenge. Cryopreservation of semen is an important potential tool to maintain genetic diversity by preserving current genetic material for future use, as well as facilitating the transport and exchange of genetic material. This study aimed to develop a protocol for semen collection and cryopreservation in the bonobo. Semen was collected from four healthy adult bonobos under general anesthesia during management translocation procedures. Semen collection utilizing urethral catheterization was not successful (n = 1), however, all males (n = 4) responded well to rectal probe electro-ejaculation. Immediately after collection, ejaculates were evaluated for color and admixtures, volume, motility, and concentration. Eosin-Nigrosin staining was prepared to evaluate morphology and viability. Ejaculates were split into two equal volumes and cryopreserved in two different extenders, using a one-step and a two-step approach. Ejaculates were gradually cooled to 4 °C in two hours, subsequently stored in liquid nitrogen vapor for twenty minutes (0.25 ml straws), and finally dropped into liquid nitrogen. RESULTS: Pre-freeze evaluation showed thick, white samples with an average ejaculate volume of 450 µl (100-1000 µl), total motility of 59% (40-80%), viability of 69% (38-85%) and 58% (46-72%) normal spermatozoa. Mainly head (22%) and tail (19%) defects were detected on the Eosin-Nigrosin stain. Ejaculates were highly concentrated, nevertheless, due to the coagulum that caused high viscosity and non-homogenous fractions, only estimations of concentration could be made (1000 million/ml). After 24 h of storage, the post-thaw evaluation showed a loss of quality with an average post-thaw total motility of 15% (5-25%) using the one-step freezing medium, and 19% (5-30%) using the two-step medium. Average post-thaw viability was 15% (4-24%) and 21% (15-29%), respectively. CONCLUSIONS: This report on ejaculates from bonobos obtained by rectal probe electro-ejaculation shows that semen parameters of this species are not completely similar to those of its sibling species, the chimpanzee. Further studies are necessary to develop an optimal protocol for the processing and cryopreservation of bonobo spermatozoa.