Abstract The cryopreservation of jaguar semen must be improved to produce high-quality biobanking doses. Until now, the rare studies of semen freezing in the species have only evaluated glycerol, always with a significant reduction in sperm quality in thawed semen. The purpose of this study was to assess the efficacy of three cryoprotectants, dimethylsulfoxide (DMSO), glycerol (GLY), and methanol (MET), in the cryopreservation of jaguar semen in an LDL-based extender, as well as the effect of thawing temperature on dosage quality. Five mature males with a history of reproduction were used. On the males, an infrared thermal image (IRT) was captured, the spicules and testes were analyzed, and the CASA system was used to evaluate the quality of fresh and thawed sperm. The superficial IRT was 4.6 ± 1.2 °C cooler than the anal sphincter, and the semen measured between 27.3 and 28.7 °C shortly after exiting the urethra. The total motility of fresh sperm was 55.3 ± 22.6%, and progressive motility was 36.3 ± 18%. The total motility of thawed sperm was 5.28 ± 2.51%, 4.49 ± %2.49, and 0.51 ± 0.62% for DMSO, GLY, and MET, respectively. DMSO and GLY performed better than MET, and there was no difference in thawing temperature (37°C 30 s vs. 50°C 12 s). All animals exhibit a considerable level of morphological changes in sperm. Low amounts of total and progressive motility were found in the thawed sperm. Males with a high level of sperm morphological changes were found to be fertile, but the lone male with normospermia was infertile. Thus, we contest the applicability of the commonly used morphological classification for bovines to felid species. highquality doses now cryoprotectants , (DMSO) (GLY) MET (MET) LDLbased LDL based extender (IRT captured analyzed 46 4 6 4. 1 2 1. C sphincter 273 27 3 27. 287 28 7 28. urethra 553 55 55. 226 22 22.6% 363 36 36. 18 18% 528 5 5.2 251 51 2.51% 449 49 4.4 249 %2.49 051 0 0.5 062 62 0.62 respectively 37°C 37C 37 vs 50C 50 s. . s) fertile infertile Thus (DMSO (GLY (MET 22.6 52 5. 25 2.51 44 24 %2.4 05 0. 06 0.6 22. 2.5 %2. 2. %2 %

Abstract The in situ population of jaguars in the Caatinga is less than 250 individuals, subdivided into five subpopulations, and is classified as endangered regarding its risk of extinction. Luisa, a 15-year-old female weighing 36 kg, was the last known ex situ jaguar from this biome. Her reproductive evaluation is detailed in this manuscript. Luisa was subjected to both a clinical and laparoscopic evaluation of her reproductive system. After 45 days of reproductive investigation, she died unexpectedly, and skin fragments were taken to establish the postmortem fibroblast lineage. At the clinical evaluation, Luisa had small, undeveloped mammary gland and a small vulva, characteristic of a nulliparous female, with no mammary gland nodules, edema, or abnormal masses. By laparoscopy, normal-appearing bladder and bowel loops were observed, as were uterine horns with standard color, shape, and length with no striae. Ovaries and uterine horns seem free of fibrinous adhesions. Both ovaries showed a yellowish color, a fibrous consistency, a decreased size (atrophied), and no follicles, hemorrhagic corpus, corpus luteum, luteal scars, or other abnormal structures. We may assume that this jaguar female was infertile based on Luisa's mature age and the absence of birthing or ovarian activity signs. The harsh conditions of the Caatinga biome, which included low food availability and frequent conflicts with humans, may have impacted both the pregnancy and lactation of Luisa’s mother and her development after birth. 25 individuals subpopulations extinction 15yearold yearold 15 year old 3 kg biome manuscript system 4 investigation unexpectedly lineage vulva nodules edema masses laparoscopy normalappearing normal appearing observed color shape striae adhesions consistency atrophied, atrophied , (atrophied) follicles luteum scars structures Luisas s signs humans birth 2 1 (atrophied

Abstract Among the different methods used for semen collection from domestic cats, the pharmacological collection by urethral catheterization becomes disruptive. Medetomidine is the elected α2-adrenoceptor agonist for that, but in several countries, it is not commercially available. This study aimed to evaluate the efficacy of detomidine compared to medetomidine in collecting semen by urethral catheterization in domestic cats. Urethral catheterization was performed on 13 mongrel cats using a disposable semi-rigid tomcat urinary catheter. Of the 19 semen collections performed with medetomidine induction, 94.7% were successful, while with detomidine induction, only 56.3% of 16 were successful. The values semen samples variables were as follows for volume - 10.56 ± 0.4 vs 8.88 ± 0.5 mL, motility - 171.67 ± 0.79 vs 49.77 ± 3.45%, vigor – 4.1 ± 0.03 vs 3.10 ± 0.1 and concentration - 3.24 ± 0.19 vs 2.15 ± 0.13 ×109 sperm/mL respectively for medetomidine and detomidine group. The failure in semen collections with detomidine was mainly due to azoospermic samples, poor urethral relaxation, insufficient volume, or contamination of urine. The sperm concentration was also lower in the detomidine group (P <0.05) when compared to medetomidine. However, when the volume of semen collected was compared, we found no statistical differences. Despite its low performance in collecting semen from cats, detomidine may be an alternative when medetomidine is not accessible.

Abstract Cryptorchidism is a genital alteration wherein one or both testicles fail to descend into the scrotum and has multifactorial causes. A free-range adult male was captured twice in the Pantanal of Nhecolândia to put a GPS collar and semen collection. Pharmacological semen collection, andrological examination and semen analysis were performed. At the first capture and during the andrological examination only the left testis was found, and the male qualified as cryptorchid. The penis had no penile spines at either procedure. The semen volume obtained at first and second capture was 435 and 160 μL, respectively, with a concentration of 618 and 100 x 106 sperm/mL, progressive motility of ~ 5% and ~ 1% and total morphological sperm abnormalities of 74% and 86%. The male was monitored by a GPS collar, but the signal was lost, making it difficult to re-captures and perform new seminal and ultrasound evaluations to discard monorchidism – exceedingly rare in felids. Genetic studies to assess the individual's homozygosity are necessary to verify whether cryptorchidism in this individual has a genetic factor.

O desenvolvimento de biotécnicas de reprodução é uma importante ferramenta para a conservação de animais silvestres ameaçados de extinção. Procedimentos de reprodução assistida em suçuarana, no entanto, são escassos na literatura, em especial aqueles relacionados à criopreservação de sêmen. Neste sentido, o presente trabalho objetivou avaliar a congelabilidade do sêmen de suçuaranas adultas mantidas em cativeiro, por meio da comparação entre duas concentrações de glicerol no meio de congelamento. Foram usados cinco machos adultos de suçuarana, mantidos no Centro de Reabilitação de Animais Silvestres do Mato Grosso do Sul (CRAS/MS). As amostras foram coletadas por eletroejaculação e avaliadas quanto ao seu aspecto físico, volume, vigor, motilidade, concentração e índice espermático. De cada ejaculado duas alíquotas foram diluídas em meio Tris-citrato-gema de ovo, em concentrações finais de 5 e 7,5% de glicerol, resfriadas a uma taxa de -0,55ºC/min e congeladas a uma taxa de -5,8ºC/min. Depois de descongeladas, as amostras foram reavaliadas e submetidas aos testes de termorresistência e hiposmótico. O protocolo de criopreservação e descongelamento de sêmen proposto se mostrou eficiente em ambas as concentrações de glicerol testadas, não havendo diferença (p>0,05) entre estas.

The development of biotechnologies of reproduction is an important tool for the conservation of wild animals threatened with extinction. Assisted reproduction procedures in mountain lions, however, are scarce, especially those related to sperm cryopreservation. In this context, this study aimed to evaluate the freezing capacity of semen from adult mountain lions in captivity through the comparison of two concentrations of glycerol in the freezing media. Five adult male mountain lions were used, held at the Rehabilitation Center for Wild Animals of Mato Grosso do Sul (CRAS/MS). Samples were collected by electroejaculation and evaluated for physical appearance, volume, sperm progressive status, sperm motility, sperm concentration and sperm motility index. Each ejaculate was spliced into two aliquots and diluted in Tris-citrate-half egg yolk, at final concentrations of 5 and 7.5% glycerol, cooled at a rate of -0.55ºC/min and frozen at a rate of -5.8ºC/min. Once thawed, the samples were re-evaluated and tested for thermoresistance and hypoosmotic swelling. The suggested protocol for cryopreservation and thawing of semen is efficient in both glycerol concentrations tested, with no difference (p>0.05) between them.