Florida manatees are semisocial marine mammals that vocalize when interacting with conspecifics and to maintain contact with offspring. While many aspects of their biology have been studied, there is a dearth of information on the diversity and complexity of vocal behavior during social, nonsocial, and stressful situations. Investigations of vocal communication repertoires which define, categorize, and correlate varied call types with behavior are needed in order to understand the behavioral and social function of associated calls. Arguably the most important social bond in manatees is the period of cow/calf dependency and empirical evidence indicates cows recognize the vocalizations of offspring. Exploration of individually distinctive vocal features can provide insight on which parameters might be salient to facilitate recognition between cows/calves. This study is focused on vocal communication in Florida manatees, how calls are structured, utilized and function while animals are distressed and during social interactions in their shallow water habitats. Hydrophones recorded vocalizations from individual calves and manatees in different behavioral contexts and varying size aggregations. Analysis of the vocal repertoire indicated manatee vocalizations can be parsed into five broadly defined call types which include the hill-shaped high squeak, tonal squeak, noisy squeal, two toned chirp, and the combinatorial squeak-squeal. Furthermore, the high squeak is likely a discrete call whereas the others are graded and do not have strict boundaries between call types (Chapter 2). Broadly defined call types were used to explore call usage with variations in behavior, group size, and group composition (Chapter 3). Manatees vocalized using few call types and altered structural parameters depending on behavioral state. Calls were longer and more frequency modulated when stressed. Vocalizations produced while cavorting were higher in entropy and more frequency modulated than when manatees were resting or feeding. Vocalizations obtained from individual calves suggest that the high squeak is a stereotypical call that is produced by smaller calves. All calves had individually distinctive acoustic features that could potentially be used in recognition (Chapter 4). Lower fundamental frequencies and higher emphasized frequencies from smaller calves suggest that the fundamental frequency may not be a reliable indicator of body size in calves. This research increases our knowledge of the vocal behavior and call characteristics of the Florida manatee. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2020. / FAU Electronic Theses and Dissertations Collection

The purpose of this study was to determine how manatees use Swallow Caye, Drowned Cayes, and Gallows Reef, three distinct habitat types within the Belize Barrier Reef lagoon system near Belize City. Data were collected using boat-based point scan methods with the assistance of volunteers. Presence/absence and photo ID methods ensured consistency of data collection despite a changing pool of volunteer researchers. Results confirmed the mangrove and seagrass ecosystem between the Belize Barrier Reef and Belize City as important manatee habitat. Inconsistent with the prevailing "seasonal distribution hypothesis" for manatees in Belize, the probability of encountering manatees at Swallow Caye and in the Drowned Cayes was equal between dry and wet seasons. However, manatees were only observed at Gallows Reef during the wet season. Swallow Caye had the highest probability of encountering manatees, confirming traditional knowledge held by local tour operators, which led to the establishment of Swallow Caye Wildlife Sanctuary in 2002. In contrast to previous studies, my data suggest that at least 44% of the manatee population carry scars from non-lethal boat strikes. The proportion of scarred animals did not vary as a function of habitat type, season, or year. The probability of encountering manatees did not change between years, despite an exponential increase in cruise ship tourism. Marine Protected Areas (MPAs) and wildlife protection laws indicate that Belize is working to meet obligations under international agreements. However, there is still cause for concern. Manatees do not remain inside designated refuge boundaries; governmental agencies depend on co-management agreements with local non-governmental organizations for enforcement of rules inside MPAs; regulations governing human behavior outside MPAs are lacking; funds for monitoring and evaluation of MPAs are lacking. Manatee conservation strategies should be integrated into a system of riverine, coastal, and marine protected areas supported by additional tactics such as required manatee training for boat captains, slow zones at hot spots outside MPAs, and continued educational outreach. With few modifications and increased enforcement and monitoring, the Belize model for manatee conservation could lead to a shared "triumph on the commons" for the manatees and the user groups that shared their habitat.

Environmental and anthropogenic stressors can affect wildlife populations in a number of ways. For marine mammals (e.g. the Florida manatee Trichechus manatus latirostris), certain stressors or conservation risk factors have been identified, but sublethal effects have been very difficult to assess using traditional methods. The development of 'biomarkers' allows us to correlate effects, such as impaired reproduction, with possible causes. A recently developed biomarker (anti-Müllerian hormone, AMH) provides an enzyme-linked immunosorbent assay of gonadal function. The study objective was to determine AMH levels in wild manatees. In total, 28 male and 17 female manatee serum samples were assayed. Animal demographics included collection date, body weight (kg) and total length (cm). In certain cases, age of individuals was also known. AMH levels ranged from 160 to 2451.85 ng ml -1 (mean = 844.65 ng ml -1) in males and 0.00 to 0.38 ng ml -1 (mean = 0.10 ng ml -1) in females. Linear regression analyses revealed a significant relationship between male AMH levels and body weight (R 2 = 0.452; p < 0.001) and length (R 2 = 0.338; p < 0.001). Due to the small sample size, regression analyses for female AMH and body weight and length were not significant. This represents the first report of AMH detection in a marine mammal. AMH levels in male manatees are the highest of any species observed to date, whereas levels in females are within reported ranges. Further studies will promote improved conservation decision by assessing AMH levels in the manatee as a function of various stressors including, but not limited to, nutritional status, serious injuries (e.g. watercraft collisions), exposure to biotoxins or contaminants, or disease.

The Florida manatee (Trichechus manatus latirostris) is a protected species that is vulnerable to both anthropogenic and natural causes of mortality. The ability of wildlife managers to oversee regulation of this species is based on available abundance estimates and mortality data. Using existing manatee mortality data collected by Florida Fish and Wildlife Conservation Commission (FWC) from 1974-2012, this study focuses on identifying significant spatial clusters of high values or “hotspots” of manatee mortality and the temporal patterns of these hotspots using the novel “emerging hotspot analysis” ArcGIS tool. The categories of manatee mortality included in this analysis were watercraft-related, perinatal, cold-stress, and other natural (which includes red tide) and were classified into five hotspot pattern categories. Of interest were the locations where consecutive or new hotspot patterns were identified among the four categories of manatee mortality included in this analysis. Consecutive hotspot clusters were found near Tampa Bay (which includes parts of Pinellas, Hillsborough, and Manatee Counties) and in the counties of Hernando/Pasco, Monroe, Palm Beach/Broward/Miami-Dade, St. Johns/Flagler, and Citrus. New hotspot clusters were found in Tampa Bay (which includes parts of Pinellas, Hillsborough, and Manatee Counties) and in the counties of Nassau, Wakulla, Charlotte/Lee, St. Lucie/Martin, Levy, Duval, Dixie, Volusia/Seminole, and Citrus. These mortality hotspots frequently overlapped areas of higher manatee and human population densities. These hotspot clusters indicate emerging patterns that highlight areas to focus future research by wildlife managers; specifically, on the relationship between human population density and concentration of watercraft activities in coastal areas, as well as the influence coastal development has on the vital resources utilized by manatees.

The distribution of organisms across the globe forms non-random patterns that are based on their selection for certain environmental features. Habitat selection is the study of how organisms choose where they live. It is a fundamental behavior shaping several ecological and evolutionary processes. The hierarchical nature of the environment makes it imperative to consider multiple scales in habitat selection studies. Even though there is extensive coverage of scaling in habitat selection literature, most published papers only consider one scale. To our knowledge, habitat selection studies of the Antillean manatee (Trichechus manatus manatus) have never included more than one spatial scale. Hence, we modeled Antillean manatee habitat selection in the Sian Ka'an Biosphere Reserve, in Mexico, at two spatial scales: study area and 1 km buffer. We used GPS coordinates of opportunistic encounters (n = 102) recorded since 2009. We randomly generated 500 pseudo-absences per presence point for both scales, and extracted the environmental conditions from each point: seagrass abundance (leaf area index or LAI), water depth, and shortest distance to land, closest river and closest resting hole. We fitted a binomial regression of the probability of presence as a function of the environmental parameters using a Bayesian approach. Our results show that the probability of manatee presence increases in proximity to resting holes for both scales and increases with land proximity at the large-scale. Overall, we showed that proximity to resting holes is the most important feature of habitat selection for Antillean manatees in Sian Ka'an, followed by proximity to land. This study demonstrates the importance of multi-scale designs in habitat selection and highlights the need for more studies looking at the use and ecological implications of manatee resting holes.

<p>Antillean manatees (Trichechus manatus manatus) were heavily hunted in the past throughout the Wider Caribbean Region (WCR), and are currently listed as endangered on the IUCN Red List of Threatened Species. In most WCR countries, including Haiti and the Dominican Republic, remaining manatee populations are believed to be small and declining, but current information is needed on their status, distribution, and local threats to the species.</p><p>To assess the past and current distribution and conservation status of the Antillean manatee in Hispaniola, I conducted a systematic review of documentary archives dating from the pre-Columbian era to 2013. I then surveyed more than 670 artisanal fishers from Haiti and the Dominican Republic in 2013-2014 using a standardized questionnaire. Finally, to identify important areas for manatees in the Dominican Republic, I developed a country-wide ensemble model of manatee distribution, and compared modeled hotspots with those identified by fishers.</p><p>Manatees were historically abundant in Hispaniola, but were hunted for their meat and became relatively rare by the end of the 19th century. The use of manatee body parts diversified with time to include their oil, skin, and bones. Traditional uses for folk medicine and handcrafts persist today in coastal communities in the Dominican Republic. Most threats to Antillean manatees in Hispaniola are anthropogenic in nature, and most mortality is caused by fisheries. I estimated a minimum island-wide annual mortality of approximately 20 animals. To understand the impact of this level of mortality, and to provide a baseline for measuring the success of future conservation actions, the Dominican Republic and Haiti should work together to obtain a reliable estimate of the current population size of manatees in Hispaniola.</p><p> </p><p>In Haiti, the survey of fishers showed a wider distribution range of the species than suggested by the documentary archive review: fishers reported recent manatee sightings in seven of nine coastal departments, and three manatee hotspot areas were identified in the north, central, and south coasts. Thus, the contracted manatee distribution range suggested by the documentary archive review likely reflects a lack of research in Haiti. Both the review and the interviews agreed that manatees no longer occupy freshwater habitats in the country. In general, more dedicated manatee studies are needed in Haiti, employing aerial, land, or boat surveys. </p><p>In the Dominican Republic, the documentary archive review and the survey of fishers showed that manatees still occur throughout the country, and occasionally occupy freshwater habitats. Monte Cristi province in the north coast, and Barahona province in the south coast, were identified as focal areas. Sighting reports of manatees decreased from Monte Cristi eastwards to the adjacent province in the Dominican Republic, and westwards into Haiti. Along the north coast of Haiti, the number of manatee sighting and capture reports decreased with increasing distance to Monte Cristi province. There was good agreement among the modeled manatee hotspots, hotspots identified by fishers, and hotspots identified during previous dedicated manatee studies. The concordance of these results suggests that the distribution and patterns of habitat use of manatees in the Dominican Republic have not changed dramatically in over 30 years, and that the remaining manatees exhibit some degree of site fidelity. The ensemble modeling approach used in the present study produced accurate and detailed maps of manatee distribution with minimum data requirements. This modeling strategy is replicable and readily transferable to other countries in the Caribbean or elsewhere with limited data on a species of interest.</p><p>The intrinsic value of manatees was stronger for artisanal fishers in the Dominican Republic than in Haiti, and most Dominican fishers showed a positive attitude towards manatee conservation. The Dominican Republic is an upper middle income country with a high Human Development Index. It possesses a legal framework that specifically protects manatees, and has a greater number of marine protected areas, more dedicated manatee studies, and more manatee education and awareness campaigns than Haiti. The constant presence of manatees in specific coastal segments of the Dominican Republic, the perceived decline in the number of manatee captures, and a more conservation-minded public, offer hope for manatee conservation, as non-consumptive uses of manatees become more popular. I recommend a series of conservation actions in the Dominican Republic, including: reducing risks to manatees from harmful fishing gear and watercraft at confirmed manatee hotspots; providing alternative economic alternatives for displaced fishers, and developing responsible ecotourism ventures for manatee watching; improving law enforcement to reduce fisheries-related manatee deaths, stop the illegal trade in manatee body parts, and better protect manatee habitat; and continuing education and awareness campaigns for coastal communities near manatee hotspots. </p><p>In contrast, most fishers in Haiti continue to value manatees as a source of food and income, and showed a generally negative attitude towards manatee conservation. Haiti is a low income country with a low Human Development Index. Only a single dedicated manatee study has been conducted in Haiti, and manatees are not officially protected. Positive initiatives for manatees in Haiti include: protected areas declared in 2013 and 2014 that enclose two of the manatee hotspots identified in the present study; and local organizations that are currently working on coastal and marine environmental issues, including research and education on marine mammals. Future conservation efforts for manatees in Haiti should focus on addressing poverty and providing viable economic alternatives for coastal communities. I recommend a community partnership approach for manatee conservation, paired with education and awareness campaigns to inform coastal communities about the conservation situation of manatees in Haiti, and to help change their perceived value. Haiti should also provide legal protection for manatees and their habitat.</p> / Dissertation

Neste estudo avaliou-se a função reprodutiva de fêmeas de peixe-boi da Amazônia (Trichechus inunguis) mantidas em cativeiro no Laboratório de Mamíferos Aquáticos do Instituto Nacional de Pesquisas da Amazônia (n=4) e no Centro de Preservação e Pesquisas de Mamíferos Aquáticos - Manaus Energia S.A. (n=2), por meio da extração e quantificação de progestinas e estrógenos fecais. As amostras fecais foram colhidas duas vezes por semana durante doze meses, foram liofilizadas e processadas de acordo com o protocolo adaptado de Schwarzenberger et al (1991). Realizou-se a validação dos radioimunoensaios em fase sólida, progesterona e 17&beta;-estradiol para o uso em extratos fecais de peixe-boi da Amazônia. A duração média do ciclo ovariano (&plusmn; EPM) definido por dois picos consecutivos de estrógenos fecais foi de 22,49 &plusmn; 1,26. A fase de estro teve duração média de 6,4 &plusmn; 0,45. Para verificar se as fêmeas mantidas em cativeiro apresentavam o padrão reprodutivo sazonal dos peixes-bois de vida livre sugerido por Best (1982), avaliou-se a influência das fases do ciclo hidrológico dos rios da bacia Amazônica. Não foi observada diferença estatística nas concentrações de progestinas fecais ao longo das fases do ciclo hidrológico, porém verificou-se que durante a fase de cheia apresentaram concentrações de estrógenos fecais superiores às outras fases, corroborando com os dados sugeridos por Best (1982). Com os resultados obtidos nesse estudo, pretende-se ampliar os conhecimentos sobre a fisiologia reprodutiva do peixe-boi da Amazônia, fornecendo mais uma ferramenta de estudo para auxiliar pesquisas e manejo reprodutivo, tendo como objetivo final a conservação da espécie. / This study has evaluated the reproductive function of six females of the Amazon manatee (Trichechus inunguis) kept in captivity at the Aquatic Mammals Laboratory - Instituto Nacional de Pesquisas da Amazônia (n=4) and Centro de Preservação e Pesquisa de Mamíferos Aquáticos-Manaus Energia S/A (n=2) through extraction and quantification of fecal estrogens and progestins. Fecal samples were obtained twice a week for 12 months. Validation of the solid phase radioimmunoassay for progesterone and 17&beta;-estradiol was performed for using on Amazon manatees fecal extracts. Ovarian cycle mean duration (&plusmn; SEM), defined by two consecutive peaks of fecal estrogens, was 22,49 &plusmn; 1,26 days. Mean estrous phase duration was 6,4 &plusmn; 0,45 days. To verify if the captive females showed the same seasonal reproductive pattern of wild manatees suggested by Best (1982), the influence of the hydrological cycle of the Amazon River Basin was analyzed. There was no statistical difference on the fecal progestins between phases, but during the flood phase it was verified that the estrogen fecal concentrations were increased, corroborating with Best (1982).These results show that the improvement of the non-invasive techniques for reproductive assessments increase the knowledge of reproductive physiology and is an important tool for scientific research, reproductive management, and species conservation.

O objetivo deste estudo foi verificar a viabilidade da dosagem de andrógenos em amostras de saliva, secreção lacrimal, urina e fezes de peixe-boi da Amazônia realizando um desafio hormonal. Dois peixes-bois amazônicos adultos machos (A-1 e A-2) foram submetidos a um protocolo de experimentação de doze dias (D-1 a D10). No D0 os animais receberam uma injeção intramuscular de GnRH exógeno. Amostras de saliva, secreção lacrimal, urina e fezes foram coletadas diariamente (entre 08h00 e 09h00) e mantidas a -20°C até o ensaio. As amostras de fezes foram liofilizadas, extraídas com metanol 80% e diluídas em tampão antes do radioimunoensaio (RIE). As amostras de urina sofreram hidrólise ácida e foram diluídas em soro bovino depletado. As amostras de saliva e secreção lacrimal foram dosadas sem etapa de extração, porém, o ensaio foi adaptado para aumentar a sensibilidade do teste. Os ensaios hormonais foram realizados utilizando um conjunto comercial de RIE para testosterona total. Um pico de andrógenos (mediana+2DI) somente foi observado nas amostras de saliva, urina e fezes de ambos os animais. Porém, os picos de andrógenos fecais ocorreram depois (cinco dias) dos picos de andrógenos urinários e salivares. Este intervalo está correlacionado com o longo tempo de passagem da digesta pelo trato gastrointestinal na espécie. Os picos salivares e urinários ocorreram muito próximos, provavelmente com poucas horas de intervalo. Esses resultados demonstram que as concentrações de andrógenos em amostras de saliva, urina ou fezes refletem consistentemente os eventos fisiológicos e são ferramentas de grande utilidade no monitoramento reprodutivo de peixes-bois da Amazônia. / The aim of this study was to verify the viability of androgens measurement in saliva, lacrimal secretion, urine and fecal samples of Amazonian manatee by carrying out hormonal challenge. Two adult male manatees (A-1 and A-2) were submitted to an experimentation protocol of twelve days (D-1 to D10). On D0 the animals received an intramuscular injection of GnRH-analogue. Salivary, lacrimal, urinary and fecal samples were collected daily (between 08h00 and 09h00) and frozen at -20°C until assayed. Fecal samples were lyophilized, extracted with 80% methanol and diluted in buffer before the radioimmunoassay (RIA). Urine samples underwent acid hydrolysis and diluted in depleted bovine serum. Salivary and lacrimal samples were assayed without extraction step, but, the assay was adapted to improve the sensibility. Hormonal assays were carried out with a commercial testosterone RIA kit. An androgen peak (>median+2IQR) was observed only in salivary, urinary and fecal samples of both animals. However, the fecal androgens peaks occurred later than urinary and salivary androgens peaks. These intervals are correlated with the long digesta passage time in this species. The salivary and urinary peaks were very close, probably with few hours of interval. These results show that androgens concentrations in saliva, urine or feces samples reflect reliably physiological events and are powerful tool for reproductive monitoring of Amazonian manatees.

O objetivo deste estudo foi verificar a viabilidade da dosagem de andrógenos em amostras de saliva, secreção lacrimal, urina e fezes de peixe-boi da Amazônia realizando um desafio hormonal. Dois peixes-bois amazônicos adultos machos (A-1 e A-2) foram submetidos a um protocolo de experimentação de doze dias (D-1 a D10). No D0 os animais receberam uma injeção intramuscular de GnRH exógeno. Amostras de saliva, secreção lacrimal, urina e fezes foram coletadas diariamente (entre 08h00 e 09h00) e mantidas a -20°C até o ensaio. As amostras de fezes foram liofilizadas, extraídas com metanol 80% e diluídas em tampão antes do radioimunoensaio (RIE). As amostras de urina sofreram hidrólise ácida e foram diluídas em soro bovino depletado. As amostras de saliva e secreção lacrimal foram dosadas sem etapa de extração, porém, o ensaio foi adaptado para aumentar a sensibilidade do teste. Os ensaios hormonais foram realizados utilizando um conjunto comercial de RIE para testosterona total. Um pico de andrógenos (mediana+2DI) somente foi observado nas amostras de saliva, urina e fezes de ambos os animais. Porém, os picos de andrógenos fecais ocorreram depois (cinco dias) dos picos de andrógenos urinários e salivares. Este intervalo está correlacionado com o longo tempo de passagem da digesta pelo trato gastrointestinal na espécie. Os picos salivares e urinários ocorreram muito próximos, provavelmente com poucas horas de intervalo. Esses resultados demonstram que as concentrações de andrógenos em amostras de saliva, urina ou fezes refletem consistentemente os eventos fisiológicos e são ferramentas de grande utilidade no monitoramento reprodutivo de peixes-bois da Amazônia. / The aim of this study was to verify the viability of androgens measurement in saliva, lacrimal secretion, urine and fecal samples of Amazonian manatee by carrying out hormonal challenge. Two adult male manatees (A-1 and A-2) were submitted to an experimentation protocol of twelve days (D-1 to D10). On D0 the animals received an intramuscular injection of GnRH-analogue. Salivary, lacrimal, urinary and fecal samples were collected daily (between 08h00 and 09h00) and frozen at -20°C until assayed. Fecal samples were lyophilized, extracted with 80% methanol and diluted in buffer before the radioimmunoassay (RIA). Urine samples underwent acid hydrolysis and diluted in depleted bovine serum. Salivary and lacrimal samples were assayed without extraction step, but, the assay was adapted to improve the sensibility. Hormonal assays were carried out with a commercial testosterone RIA kit. An androgen peak (>median+2IQR) was observed only in salivary, urinary and fecal samples of both animals. However, the fecal androgens peaks occurred later than urinary and salivary androgens peaks. These intervals are correlated with the long digesta passage time in this species. The salivary and urinary peaks were very close, probably with few hours of interval. These results show that androgens concentrations in saliva, urine or feces samples reflect reliably physiological events and are powerful tool for reproductive monitoring of Amazonian manatees.

Neste estudo avaliou-se a função reprodutiva de fêmeas de peixe-boi da Amazônia (Trichechus inunguis) mantidas em cativeiro no Laboratório de Mamíferos Aquáticos do Instituto Nacional de Pesquisas da Amazônia (n=4) e no Centro de Preservação e Pesquisas de Mamíferos Aquáticos - Manaus Energia S.A. (n=2), por meio da extração e quantificação de progestinas e estrógenos fecais. As amostras fecais foram colhidas duas vezes por semana durante doze meses, foram liofilizadas e processadas de acordo com o protocolo adaptado de Schwarzenberger et al (1991). Realizou-se a validação dos radioimunoensaios em fase sólida, progesterona e 17&beta;-estradiol para o uso em extratos fecais de peixe-boi da Amazônia. A duração média do ciclo ovariano (&plusmn; EPM) definido por dois picos consecutivos de estrógenos fecais foi de 22,49 &plusmn; 1,26. A fase de estro teve duração média de 6,4 &plusmn; 0,45. Para verificar se as fêmeas mantidas em cativeiro apresentavam o padrão reprodutivo sazonal dos peixes-bois de vida livre sugerido por Best (1982), avaliou-se a influência das fases do ciclo hidrológico dos rios da bacia Amazônica. Não foi observada diferença estatística nas concentrações de progestinas fecais ao longo das fases do ciclo hidrológico, porém verificou-se que durante a fase de cheia apresentaram concentrações de estrógenos fecais superiores às outras fases, corroborando com os dados sugeridos por Best (1982). Com os resultados obtidos nesse estudo, pretende-se ampliar os conhecimentos sobre a fisiologia reprodutiva do peixe-boi da Amazônia, fornecendo mais uma ferramenta de estudo para auxiliar pesquisas e manejo reprodutivo, tendo como objetivo final a conservação da espécie. / This study has evaluated the reproductive function of six females of the Amazon manatee (Trichechus inunguis) kept in captivity at the Aquatic Mammals Laboratory - Instituto Nacional de Pesquisas da Amazônia (n=4) and Centro de Preservação e Pesquisa de Mamíferos Aquáticos-Manaus Energia S/A (n=2) through extraction and quantification of fecal estrogens and progestins. Fecal samples were obtained twice a week for 12 months. Validation of the solid phase radioimmunoassay for progesterone and 17&beta;-estradiol was performed for using on Amazon manatees fecal extracts. Ovarian cycle mean duration (&plusmn; SEM), defined by two consecutive peaks of fecal estrogens, was 22,49 &plusmn; 1,26 days. Mean estrous phase duration was 6,4 &plusmn; 0,45 days. To verify if the captive females showed the same seasonal reproductive pattern of wild manatees suggested by Best (1982), the influence of the hydrological cycle of the Amazon River Basin was analyzed. There was no statistical difference on the fecal progestins between phases, but during the flood phase it was verified that the estrogen fecal concentrations were increased, corroborating with Best (1982).These results show that the improvement of the non-invasive techniques for reproductive assessments increase the knowledge of reproductive physiology and is an important tool for scientific research, reproductive management, and species conservation.