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      Elaboration of massage technique for semen collection and examination of semen characteristics in chinchilla ( Chinchilla lanigera)

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          Abstract

          The practice of artificial insemination for the long-tailed chinchilla has not been fully elaborated to date, and existing data available regarding their reproduction properties is contradictory. Until now, the collection of semen for chinchillas has been most-commonly obtained using electro-ejaculation methods exclusively. The primary objective of this study was the development of a manual technique for semen collection which meets all animal welfare requirements. An additional aim was to determine the basic spermatological parameters, such as motility, concentration, type and ratio of morphological abnormalities and live/dead cell ratio, under typical northern-hemisphere conditions, in Hungary. Over a 3 month period, a special massage technique was developed for the study, and using this method, the sperm parameters of 46 males were subsequently analyzed weekly for a period of one year. Approximately 66% of chinchillas responded positively to this technique, with the success rate of semen-collection attempts showing no variation between seasons. Average sperm concentration for the whole year was 935.17 million/ml using this method. Total cell motility was the highest in winter (90.3%), and the lowest in spring (84.3%). The proportion of live, intact cells were above 80% on average for the year, while the ratios of live, morphologically abnormal and dead cells were 6% and 14%, respectively. We found that midpiece abnormalities occurred in the highest proportion (0.95%-3.38%), while the head abnormalities showed the lowest ratio (0.01%-0.15%). Standard deviation among the parameters was relatively high, with the spring season proving to be the weakest in terms of sperm quality. This study has demonstrated that, semen can be successfully collected without the use of electro-ejaculation or anesthesia. Furthermore, although spermatological parameters do exhibit some fluctuation for the different times of the year, semen collected is nonetheless suitable for the purpose of artificial insemination of chinchillas at any time.

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          Semen collection in the dog.

          This review will discuss semen collection in the dog. Semen samples may be collected from male dogs for the purposes of artificial insemination, cryopreservation or diagnosis. The materials needed for semen collection depend on which method is used and the collector's level of expertise with this procedure. At minimum, two sterile centrifuge tubes or specimen cups can be used to collect semen as it is ejaculated (for the combined first and second fractions and for the third fraction). The most common method for semen collection in the dog is by digital stimulation. Under ideal conditions, this procedure is performed in the presence of an estrous bitch. Initially, the dog's penis is vigorously massaged through the prepuce at the level of the bulbus glandis (caudal-most aspect of the prepuce) until a partial erection develops (initial engorgement of the bulbus glandis). The prepuce is quickly retracted past the bulbus glandis and firm constant pressure is applied to the penis behind the bulbus glandis by squeezing the penis between index finger and thumb. Pelvic thrusting may occur following application of pressure behind the bulbus glandis during the development a "full" erection. The ejaculate is composed of three fractions: first (sperm-poor), second (sperm-rich) and third (prostatic fluid). In addition to digital stimulation of the penis, spermatozoa have been collected from dogs using electroejaculation and pharmacologic methods.
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            Factors affecting sperm motility. VII. Sperm viability as affected by change of pH and osmolarity of semen and urine specimens.

            The effects of pH and osmolarity of semen and urine specimens on motility and velocity of human spermatozoa were studied objectively with the aid of the multiple exposure photography (MEP) method. The pH of fresh ejaculates ranged from 7.2 to 8.2 and specimens were slightly hyperosmotic ranging between 300 to 380 mOsm/kg. Gradually changing the pH and osmolarity to either side of normal values led to progressive loss of sperm motility. However, sperm velocity was slightly increased by mild alkalinization and hyperosmolarity. Spermatozoa that became immobilized by acidification regained their motility shortly after pH was restored to normal values. In the majority of instances spermatozoa lost their motility when mixed with fresh urine specimens. Neutralization of urinary pH could not protect them from this effect unless urine osmolarity was also isotonically adjusted. It is suggested that patients with retrograde ejaculation should adequately increase their fluid intake before recovery of sperm from their bladder for artificial insemination.
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              Electroejaculation in the Chinchilla (Chinchilla lanigera): effects of anesthesia on seminal characteristics.

              Repetitive electroejaculation is probably extremely stressful in conscious animals and could adversely affect fertility. The present study was designed to (a) evaluate the effects of anesthesia (40 mg ketamine/kg body weight, i.m.) on a method of electroejaculation used previously in conscious chinchillas (Chinchilla lanigera), and (b) determine the quality of the electroejaculated semen obtained under anesthesia. In Experiment 1 (8 animals), a 4 x 4 Latin square design was used to study the effects of anesthesia and ejaculatory voltage on semen collection, ejaculate volume, sperm concentration, motility, viability, response to the hypoosmotic swelling test (HOST), and acrosomal status. In Experiment 2 (12 animals), the effects of differing voltages and the number of stimuli on ejaculation by conscious or anesthetized males was determined. In both experiments, all the conscious animals ejaculated, but only 60% ejaculated under anesthesia and they required more stimuli and higher voltages to achieve ejaculation. Ejaculate volume was significantly lower in anesthetized ( 40 microl), but sperm concentration was unaffected. None of the indices of sperm quality were affected by anesthesia. The techniques we have developed in anesthetized domestic chinchillas should be applicable to endangered chinchillas in the wild, and though the number of sperm available is reduced, there are still sufficient for assisted reproduction.
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                Author and article information

                Contributors
                Role: Data curationRole: InvestigationRole: MethodologyRole: Writing – original draft
                Role: Investigation
                Role: Supervision
                Role: Investigation
                Role: Investigation
                Role: Project administration
                Role: Funding acquisition
                Role: Investigation
                Role: InvestigationRole: SupervisionRole: Writing – original draft
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                PLOS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                31 August 2023
                2023
                : 18
                : 8
                : e0290441
                Affiliations
                [1 ] Hungarian University of Agriculture and Life Sciences, Institute of Genetics and Biotechnology, Department of Animal Biotechnology, Reproductive Bilogy and Toxicology Group, Gödöllő, Hungary
                [2 ] National Centre for Biodiversity and Gene Conservation, Institute for Farm Animal Gene Conservation, Institute for Gene Conservation Science and Small Animal Research, Gödöllő, Hungary
                Zhejiang University College of Life Sciences, CHINA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                https://orcid.org/0009-0002-3170-2569
                https://orcid.org/0000-0001-9302-1115
                Article
                PONE-D-22-30686
                10.1371/journal.pone.0290441
                10471009
                37651370
                048bd554-7879-4c30-8565-6eb77549836f
                © 2023 Babarczi et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 7 November 2022
                : 8 August 2023
                Page count
                Figures: 5, Tables: 2, Pages: 11
                Funding
                Funded by: Operational Programme of Economic Development and Innovation / Hungary
                Award ID: GINOP-2.1.7-15-2016-02232
                This study was supported by the Operational Programme of Economic Development and Innovation / Hungary (Project code: GINOP-2.1.7-15-2016-02232). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cellular Types
                Animal Cells
                Germ Cells
                Sperm
                Biology and Life Sciences
                Anatomy
                Body Fluids
                Semen
                Medicine and Health Sciences
                Anatomy
                Body Fluids
                Semen
                Biology and Life Sciences
                Physiology
                Body Fluids
                Semen
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Mammals
                Rodents
                Chinchillas
                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
                Mammals
                Rodents
                Chinchillas
                Earth Sciences
                Seasons
                Spring
                Biology and Life Sciences
                Cell Biology
                Cell Motility
                Earth Sciences
                Seasons
                Earth Sciences
                Seasons
                Winter
                Earth Sciences
                Seasons
                Autumn
                Custom metadata
                All relevant data are within the paper.

                Uncategorized
                Uncategorized

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