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Crocodylian conservation and evolution insights from an anatomical and histological examination of phalli from male false gharial (Tomistoma schlegelii)

Moore BC, Fitri WN, Augustine L

Anat Histol Embryol, 2020 May;49(3):390-401.
PMID: 32154618 DOI: 10.1111/ahe.12542

As wild population threats for the endangered false gharial (Tomistoma schlegelii) persist, conservation breeding programs, including developing semen collection techniques for subsequent artificial insemination, are becoming important species conservation measures. Developing reproductive biology understanding of a species is important to developing best practices and hopefully maximizing reproductive successes. However, information on crocodylians functional copulatory anatomy in general is lacking. To that end, zoological facilities and conservation centres have the exceptional opportunity to contribute new understandings that may not otherwise be attainable regarding crocodylian reproductive anatomy, particularly during routine physical examinations or post-mortem necropsies. Therefore, to better understand T. schlegelii reproductive biology, to contribute knowledge in support of zoo breeding conservation efforts and to contribute to what is known overall about crocodylian reproduction, we investigated phallic anatomy of adult male Tomistoma from two zoological populations, the St. Louis Zoo, USA and Sungai Dusun Wildlife Reserve, Peninsular Malaysia. Here, we present the gross anatomical features and histological analysis of underlying tissue-level details in pursuit of a better understanding of copulatory function and associated gamete transfer mechanisms. While much of the overall Tomistoma phallic morphology and inferred function corresponds to that of other crocodylian species and speaks to conserved aspects of functional anatomy across taxa, species-specific aspects of glans and glans tip morphology are also identified. These novelties are discussed in a general function and overall broader evolutionary contexts.
Fulltext The Comparison of Semen Collection in Electroejaculation, Rectal Massage and Combination of Both Methods in the Critically Endangered Malayan Pangolin, Manis javanica

Tarmizi R, Keng Chee Y, Sipangkui S, Zainuddin ZZ, Fitri WN

Animals (Basel), 2020 Oct 23;10(11).
PMID: 33113883 DOI: 10.3390/ani10111948

This article describes the semen characteristics from different collection methods between captive and confiscated Malayan pangolins, Manis javanica. Semen was collected from 15 pangolins; two captive and 13 confiscated individuals at the mean weight of 9.36 ± 1.94 kg. The three semen collection methods employed were electroejaculation, rectal massage and a combination of both techniques. The semen characteristics (mean ± standard deviation) of the Malayan pangolin are volume (73.75 ± 144.57 µL), pH (7.63 ± 0.53), spermatozoa concentration (997.19 ± 728.98 × 106 /mL), total motility (59.60% ± 30.00%), progressive motility (48.95% ± 30.93%), mass motility (3.50 ± 1.50) and live spermatozoa (80.25% ± 13.45%). There was no significant difference in semen characteristics between the three collection methods. The percentages of live spermatozoa were significantly different, suggesting better samples from captive compared to confiscated animals. However, there was no significant difference in spermatozoa kinetics between the captive and confiscated samples, suggesting the potential of utilizing confiscated individuals for gamete recovery to conserve the genetic pool of pangolins. All three methods of semen collection were successfully performed in pangolins and should be considered; however, electroejaculation remains the most consistent method of obtaining semen from the species.
Fulltext Opportunities and challenges associated with fecal progesterone metabolite analysis

Peter ID, Haron AW, Jesse FFA, Ajat M, Han MHW, Fitri WN, et al.

Vet World, 2018 Nov;11(10):1466-1472.
PMID: 30532503 DOI: 10.14202/vetworld.2018.1466-1472

Conventionally, plasma or milk progesterone evaluations are used to determine the reproductive status of female animals. Collection of such samples is often associated with difficulties of animal handling and restraint. Measurable quantities of progesterone metabolites are found in feces of animals. Their concentrations are known to be well correlated to plasma progesterone levels and are, therefore, used as non-invasive samples for assessing reproductive function in a wide range of animal species. Although the analysis of fecal progesterone metabolites has been widely accepted in many laboratories, several factors are known to affect the results from this valuable analytical technique. Some of these factors include storage/transportation media for fecal samples, type of solvent that is used for extraction of progesterone metabolites from feces, and the type and sensitivity of an assaying technique employed. Although fecal progesterone metabolites analysis is associated with some difficulties, it can effectively be used to monitor reproductive function in a wide range of animal species. This review aims to highlight the usefulness of fecal progesterone metabolite analysis as a non-invasive technique in monitoring reproductive function in animals. The article mainly focuses on the many opportunities and challenges associated with this analytical technique.
Fulltext Semen characteristics, extension, and cryopreservation of Rusa deer (Rusa timorensis)

Fitri WN, Wahid H, Rosnina Y, Jesse FFA, Aimi-Sarah ZA, Mohd-Azmi ML, et al.

Vet World, 2017 Jul;10(7):779-785.
PMID: 28831222 DOI: 10.14202/vetworld.2017.779-785

AIM: The objective of this research is to report parameters for breeding soundness evaluation, semen extension, and cryopreservation in Rusa timorensis.
MATERIALS AND METHODS: Seven healthy stags were chosen for semen collection using an electroejaculator. The collections were performed twice in a breeding season between February and June 2016. Samples were collected between 2 and 3 weeks interval, collected twice for each animal. Semen was evaluated, extended, and cryopreserved using four different extenders; Andromed®, BioXcell®, Triladyl®, and a modified Tris-egg yolk combined with Eurycoma longifolia Jack.
RESULTS: R. timorensis semen characteristics according to volume (ml), color, sperm concentration (106/ml), general motility (%), progressive motility (%), and % morphology of normal spermatozoa are 0.86±0.18 ml, thin milky to milky, 1194.2±346.1 106/ml, 82.9±2.8%, 76.1±4.8%, and 83.9±4.8%, respectively.
CONCLUSION: Semen characteristics of R. timorensis collected by electroejaculation is good allowing for cryopreservation and future artificial insemination work. The most suitable extender for Rusa deer semen is Andromed®.