Biological characteristics of pregnancy in captive Yangtze finless porpoises revealed by urinary metabolomics†.

The Yangtze finless porpoises (Neophocaena asiaeorientalis a.) are an endemic and critically endangered species in China. Intensive captive breeding is essential for understanding the biology of critically endangered species, especially their pregnancy characteristics, knowledge of which is crucial for effective breeding management. Urine metabolomics can reveal metabolic differences, arising from physiological changes across pregnancy stages. Therefore, we used the urinary metabolomic technology, to explore urinary metabolite changes in pregnant Yangtze finless porpoises. A total of 2281 metabolites were identified in all samples, which including organic acids and derivatives (24.45%), organoheterocyclic compounds (20.23%), benzenoids (18.05%), organic oxygen compounds (7.73%), and phenylpropanoids and polyketides (6.48%). There were 164, 387, and 522 metabolites demonstrating differential abundance during early pregnancy, mid pregnancy, and late pregnancy, respectively, from the levels observed in nonpregnancy. The levels of pregnenolone, 17α-hydroxyprogesterone, and tetrahydrocortisone were significantly higher during all pregnancy stages, indicating their important roles in fetal development. The differential metabolites between nonpregnancy and pregnancy were mainly associated with amino acid and carbohydrate metabolism. Moreover, metabolic activity varied across pregnancy stages; steroid hormone biosynthesis was predominant in early pregnancy, and amino acid biosynthesis and carbohydrate metabolism were predominant in mid pregnancy and late pregnancy, respectively. Our results provide new insights into metabolic characteristics in the Yangtze finless porpoises' urine during pregnancy, and indicate that the differential levels of urine metabolites can determine pregnancy in Yangtze finless porpoises, providing valuable information for the husbandry and management of pregnant Yangtze finless porpoises in captivity.

Physiological Function of Gut Microbiota and Metabolome on Successful Pregnancy and Lactation in the Captive Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis).

The gestation period in captive Yangtze finless porpoise (YFP) is a well-coordinated and dynamic process that involves both systemic and local alterations. The gut microbiota and its connection to fecal metabolites are crucial in supporting fetal development and ensuring maternal health during reproductive stages. This study evaluates changes in the gut microbiota and their correlation with fecal metabolites in captive YFPs during different reproductive stages. The results reveal that microbial community structure changed significantly during reproductive stages, while gut microbial diversity remained stable. The genus unclassified Peptostrptococcaceae, Corynebacterium, and norank KD4-96 were significantly greater in non-pregnancy (NP), Terrisporobacter was significantly greater in lactating (LL), and Clostridium was significantly higher in early-pregnancy (EP) compared to the other groups. The host fecal metabolome exhibited significant alterations during the reproductive stages. Indoxyl sulfate, octadecatrienoic acid, and methionyl-methionine were significantly higher in the NP; galactosylglycerol, chondroitin 6-sulfate, and lumichrome were significantly higher in the EP and mid-pregnancy (MP); and valylleucine and butyryl-l-carnitine were significantly higher in the LL. The altered metabolites were mostly concentrated in pathways associated with arachidonic acid metabolism (significantly altered in NP), leucine, valine, and isoleucine biosynthesis (significantly altered in EP and MP), and glycerophospholipid metabolism (significantly altered in LL compared to others stages). Additionally, we found a strong link between variations in the host metabolism and alterations in the fecal bacteria of captive YFP. In conclusion, this study provides detailed insights into host metabolic and fecal bacterial changes in captive YFP during reproduction stages, providing important knowledge for improving the reproductive management in the captive YFP.

Proteus faecis: a potentially pathogenic bacterium isolated from the freshwater Yangtze finless porpoise.

Proteus faecis is a gram-negative facultative anaerobic rod-shaped bacterium capable of swarming motility. It has been isolated from numerous sources such as humans, animals, and refuse and is considered potentially pathogenic towards humans. In this study, bacteria were isolated from the blowhole of a Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP) living in captivity in China. One bacterium, P. faecis porpoise, was isolated and whole genome sequencing done. Biofilm formation, motility and antimicrobial resistance were also investigated. To find putative virulence factors, the genome of P. faecis strain porpoise was compared to the genomic sequences of eight other P. faecis isolates using the Bacterial and Viral Bioinformatics Resource Center (BV-BRC) ( https://www.bv-brc.org/ ). The goal of this study was to initially characterize the pathogenicity of this bacterium isolated from a cetacean species using both pathogenomics and conventional approaches.

Biofilm formation, antimicrobial assay, and toxin-genotypes of Clostridium perfringens type C isolates cultured from a neonatal Yangtze finless porpoise.

This is a culture-dependent study with the objective of pure culturing and characterizing pathogenic bacteria from the blowhole, lung, stomach and fecal samples of a neonatal crucially endangered Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis) that died 27 days after birth. Bacteria were inoculated using a swab onto blood and MacConkey agar plates and representative isolates were identified through 16S rRNA gene sequence analysis. A total of three Clostridium perfringens type C strains from the fecal samples were isolated. Toxin genes, including cpa, cpb and cpb2, were detected by PCR amplification, whereas the etx, iap and cpe genes were not detected. Biofilm formation of the three strains was then examined. Only one strain was capable of biofilm formation. In addition, isolates showed strong resistance against the antibiotics amikacin (3/3), erythromycin (1/3), gentamicin (3/3), streptomycin (3/3), and trimethoprim (3/3), while sensitivity to ampicillin (3/3), bacitracin (3/3), erythromycin (2/3), penicillin G (3/3), and tetracycline (3/3). The results suggested C. perfringens type C could have contributed to the death of this neonatal porpoise.

Contextual impacts on individual and synchronous breathing rate variations in three captive odontocete groups.

Breathing rates are often collected both in the wild and in captivity to inform on cetaceans' internal state. However, few studies have investigated the effect of various factors on this breathing rate. We investigated the variations of individual and synchronous breathing rates depending on individual features (species, sex, age), displayed behavior, social parameters (social grouping), and environmental parameters (diurnal variation, presence of enrichment, unusual events, and presence of visitors in three groups of captive odontocetes (Yangtze finless porpoises, Neophocaena asiaeorentalis asiaeorientalis, East-Asian finless porpoises, Neophocaena asiaeorentalis sunameri, bottlenose dolphins, Tursiops truncatus). Both individual and synchronous breathing rates were the highest when animals engaged in energetic or social behaviors. Individual breathing rate decreased but synchronous breathing rate increased with the presence of enrichment. Both rates increased during unusual events (e.g., pool cleaning, presence of a diver in the pool, noise, transport) and when public was present for Yangtze finless porpoises. Finally, synchronous breathing rate increased for Yangtze finless porpoises when experiencing social separation. We suggest that individual and synchronous breathing rates are useful parameters to measure, both in wild and captive animals, to obtain information on their arousal/stress state. However, these rates should be interpreted with caution and should be used together with other parameters to allow more accurate inferences.

Putative virulence factors of Plesiomonas shigelloides.

Plesiomonas shigelloides is a Gram-negative rod-shaped bacterium which has been isolated from humans, animals and the environment. It has been associated with diarrhoeal disease in humans and various epizootic diseases in animals. In this study P. shigelloides strains were isolated from the faecal material of a captive Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP) living in semi-natural conditions in China. Plesiomonas shigelloides strain EE2 was subjected to whole genome sequencing. The draft genome was then compared to the genome sequences of ten other P. shigelloides isolates using the Pathosystems Resource Integration Center pipeline. In addition to several virulence factors which have been previously reported, we are proposing new candidate virulence factors such as a repeats-in-toxin protein, lysophospholipase, a twin-arginine translocation system and the type VI secretion effector Phospholipase A1.

Isolation of culturable aerobic bacteria and evidence of Kerstersia gyiorum from the blowhole of captive Yangtze finless porpoises.

Bacterial respiratory illnesses are problematic in aquatic mammals such as the Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP), which is now at a critically endangered status. Yet little is known about the bacteria inhabiting the respiratory tract of YFPs. In this study, we preliminarily characterized the culturable aerobic bacteria from blow samples of captive YFPs. The bacterial diversity was assessed through cultivation by direct exhalation onto Columbia blood agar plates and identification of representative isolates through 16S rRNA gene sequence analysis. In total, eleven bacterial species belonging to four phyla Proteobacteria (71 %), Firmicutes (25 %), Bacteroidetes (3 %) and Actinobacteria (1 %) were identified. Most of these isolates were opportunistic pathogens found in respiratory illnesses in humans and animals. We also reported the first case of Kerstersia gyiorum isolated from an animal. This work provides a preliminary assessment of the bacteria present in the respiratory tract of captive YFPs, which will be an important first step in elucidating the roles of normal microbiota in maintaining respiratory health of YFPs. This study also points out the necessity of future long-term monitoring of blowhole microorganisms in the YFPs and making emergency preparedness plans for respiratory tract infections. These measures can aid in assessing the pathogenic risk of the critically endangered YFP populations.

Fecal Bacterial Composition of the Endangered Yangtze Finless Porpoises Living Under Captive and Semi-natural Conditions.

Intestinal microbiota is essential to the health and physiology of host animals. We undertook the first microbiological study of the fecal bacterial composition from critically endangered (CR) Yangtze finless porpoises (Neophocaena asiaeorientalis asiaeorientalis; YFPs) living under captive and semi-natural conditions using both high-throughput sequencing method and 16S rRNA gene clone library method. As determined by high-throughput sequencing of V3-V4 regions of the 16S rRNA gene, semi-natural samples harbored 30 and 36 operational taxonomic units (OTUs), which was more than the 22 and 27 OTUs detected from YFPs living in captivity. In captive YFPs Firmicutes was the predominant phylum, whereas this was Proteobacteria for YFPs living in semi-nature conditions. This suggests habitat-specific fecal bacterial composition of YFPs. Plesiomonas spp. and Aeromonas spp., which are potentially pathogenic, were identified in all the feces. Bacterial diversity from one porpoise living in captivity was also determined by constructing a 16S rRNA gene clone library and only 1 phylum was identified. High-throughput sequencing was more effective at determining the bacterial diversity compared to the 16S rRNA gene clone library. This study provides important information for the management and conservation of the CR YFPs.

Parentage-Based Group Composition and Dispersal Pattern Studies of the Yangtze Finless Porpoise Population in Poyang Lake.

Social behaviors are poorly known for the critically endangered Yangtze finless porpoise (YFP, Neophocaena asiaeorientalis asiaeorientalis). Here, group composition and dispersal patterns of the YFP population living in the Poyang Lake were studied by parentage-based pedigree analyses using 21 microsatellite loci and a 597 bp segment of the mitochondrial DNA control region. In this study, 21 potential mother-offspring pairs and six potential father-offspring pairs (including two potential parents-offspring pairs) were determined, among which 12 natural mother-offspring groups and a maternal group of three generations were found. No genetically-determined fathers were found associated with their offspring. This study also found that maternally related porpoises at the reproductive state tend to group together. This suggest maternal relationship and reproductive state may be factors for grouping in the YFP population. In natural mother-offspring groups, male offspring were all younger than two years old, which suggest male offspring may leave their mothers at approximately two years of age, or at least they were not in tight association with their mothers as they may have been under two years old. However, female offspring can stay longer with their mothers and can reproduce in the natal group.

De novo assembly and characterization of narrow-ridged finless porpoise renal transcriptome and identification of candidate genes involved in osmoregulation.

During the evolutionary transition from land to water, cetaceans have undergone numerous critical challenges, with osmoregulation being the major one. Two subspecies of the narrow-ridged finless porpoise (Neophocaena asiaeorientalis), the freshwater Yangtze finless porpoise (N. a. asiaeorientalis, NAA) and the marine East Asian finless porpoise (N. a. sunameri, NAS), provide excellent subjects to understand the genetic basis of osmoregulatory divergence between freshwater and marine mammals. The kidney plays an important and well-established role in osmoregulation in marine mammals and thus, herein, we utilized RNA-seq to characterize the renal transcriptome and preliminarily analyze the divergence between the NAA and the NAS. Approximately 48.98 million clean reads from NAS and 49.40 million clean reads from NAA were obtained by RNA-Seq. And 73,449 (NAS) and 68,073 (NAA) unigenes were assembled. Among these annotations, 22,231 (NAS) and 21,849 (NAA) unigenes were annotated against the NCBI nr protein database. The ion channel complex GO term and four pathways were detected as relevant to osmoregulation by GO and KEGG pathway classification of these annotated unigenes. Although the endangered status of the study species prevented analysis of biological replicates, we identified nine differentially expressed genes (DEGs) that may be vital in the osmoregulation of the narrow-ridged finless porpoise and worthwhile for future studies. Of these DEGs, the differential expression and distribution of the aquaporin-2 (AQP2) in the collecting duct were verified using immunohistochemical experiments. Together, this work is the first report of renal transcriptome sequencing in cetaceans, and it will provide a valuable resource for future molecular genetics studies on cetacean osmoregulation.

Spatial distribution, trophic magnification, and risk assessment of per- and polyfluoroalkyl substances in Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis): Risks of emerging alternatives.

The Yangtze finless porpoise (YFP, Neophocaena asiaeorientalis asiaeorientalis) is the only freshwater cetacean found in China. However, per- and polyfluoroalkyl substances (PFASs) risks in YFPs remain unclear. In this study, legacy PFASs, their precursors and alternatives, were determined in YFP muscles (n = 32), liver (n = 29), kidney (n = 24), skin (n = 5), and blubbers (n = 25) collected from Poyang Lake (PL) and Yangtze River (YR) between 2017 and 2023. Perfluorooctane sulfonic acid (PFOS) was the predominant PFAS in all YFP tissues, with a median hepatic concentration of 1700 ng/g wet weight, which is higher than that in other finless porpoises worldwide. PFOS, chlorinated polyfluorinated ether sulfonates (Cl-PFESAs), and perfluoroalkane sulfonamides concentrations in YFP livers from PL were significantly higher than those from YR (p < 0.05); however, the opposite was observed for hexafluoropropylene oxide acids. Biomagnification and trophic magnification factors (BMF and TMF, respectively) of most PFASs in the YFP food web were > 1. Perfluoroheptane sulfonic acid had the highest BMF value (99), followed by 6:2 Cl-PFESA (94) and PFOS (81). The TMFmuscle and TMFliver values of the total PFASs were 3.4 and 6.6, respectively, and were significantly positively correlated with the fluorinated carbon chain length (p < 0.01). In addition, up to 62 % of the hazard quotients for 6:2 Cl-PFESA were > 1, which was higher than that of PFOS (48 %), suggesting a high hepatotoxicity of 6:2 Cl-PFESA to YFPs. Bioaccumulation and biotoxicity of legacy and emerging alternatives in aquatic organisms continue to be a concern, especially for underscoring the vulnerability of the long-lived and endangered species.

Cytotoxicity and mechanisms of perfluorobutane sulfonate (PFBS) in umbilical cord fibroblast cells of Yangtze finless porpoise.

Yangtze finless porpoises (YFP) accumulate high levels of per- and polyfluoroalkyl substances (PFASs). However, the health impacts of PFASs to YFP are still unknown because it is technically and ethically unfeasible to use the critically endangered YFP in toxicological exposures. To uncover the potential toxicities of PFASs to YFP, this study exposed a YFP umbilical cord fibroblast cell line to perfluorobutane sulfonate (PFBS), an emerging PFASs pollutant in the aquatic environments. After exposure, the cytotoxicity and mechanisms of PFBS were explored. Our preliminary experiments found that PFBS compromised the cell viability in a concentration and duration dependent manner. In an exposure of 48-h duration, the maximum no observed effect concentration (NOEC) of PFBS was determined to be 400 µM. High-throughput proteomics were then conducted to identify the differentially expressed proteins in YFP cells exposed to 400 µM PFBS for 48 h. The results found that PFBS exposure significantly perturbed the proteome fingerprints of YFP umbilical cord fibroblast cells. Functional annotation of differential proteins showed that PFBS had the potential to impair a variety of biological processes associated with the immunity, oxidative stress, metabolism, and proteolysis. Consistently, the intracellular levels of reactive oxygen species (ROS) and proinflammatory cytokine IL-1ß were significantly increased by PFBS in YFP umbilical cord fibroblast cells. Overall, this study highlights the toxic effects of emerging PFASs on YFP and provides reference data to evaluate the health risks of aquatic pollution under the context of national YFP protection. To our knowledge, this is the first omics study using YFP umbilical cord fibroblast cells in ecotoxicology of PFASs, which is applicable to various cetacean species and pollutants.

Accumulation and risk prioritization of psychoactive substances in the critically endangered Yangtze finless porpoise.

Psychoactive substances have been identified as a kind of emerging contaminants in aquatic environment and pose potential adverse effects on aquatic animals. Yangtze finless porpoise, a critically endangered species in China, is also facing the threat of psychoactive substances. In this study, the accumulation characteristics and risk prioritization of psychoactive substances were investigated in Yangtze finless porpoise collected from Poyang Lake (PYL) and Tian-E-Zhou Oxbow (TZO) in Yangtze River basin. The levels of psychoactive substances were detected in the range of below method detection limits (MDLs) to 98.4 ng/mL in the serum of Yangtze finless porpoise. Codeine (COD) and methamphetamine were identified as the major substances due to the highest residual levels with a median concentration of 0.72 ng/mL and 0.33 ng/mL, respectively. The total concentrations of psychoactive substances in the porpoise collected from TZO was significantly higher than those from PYL. Risk analysis based on effect ratio derived from the ratio of steady-state psychoactive substance serum concentration in the porpoise and human therapeutic plasma concentration revealed that COD was the substance with the highest risk among the psychoactive substances detected, followed by lysergic acid diethylamide (LSD), morphine, alprazolam (ALPZ) and lormetazepam. Location-specific risk prioritization of psychoactive substances found that the top 3 substances are LSD, lorazepam (LORZ) and ALPZ in PYL, and COD, LSD and LORZ in TZO. The results disclose the accumulation of psychoactive substances in Yangtze finless porpoise and suggest that the potential adverse effects should be concerned.

Viral Diversity and Epidemiology in Critically Endangered Yangtze Finless Porpoises (Neophocaena asiaeorientalis asiaeorientalis).

The Yangtze finless porpoise (YFP) (Neophocaena asiaeorientalis asiaeorientalis) is a critically endangered freshwater cetacean, with about 1,249 individuals thought to be left in the wild. However, viral entities and viral diseases of YFPs remain obscure. In this study, anal swabs for virome analysis were collected during the physical examination of YFPs in the Tian-E-Zhou Oxbow (TEO) ex situ reserve. A total of 19 eukaryotic viral species belonging to 9 families, including Papillomaviridae, Herpesviridae, Picornaviridae, Picobirnaviridae, Caliciviridae, Retroviridae, Parvoviridae, Virgaviridae, and Narnaviridae, and other unclassified viruses were identified based on metasequencing. Among these detected viruses, a novel herpesvirus (NaHV), two different kobuviruses (NaKV1-2), and six different papillomaviruses (NaPV1 to -6) were considered potential risks to YFPs and confirmed by PCR or reverse transcription-PCR (RT-PCR). Most YFPs sampled were found to harbor one or more kinds of detected viral genomes (52/58 [89.7%]). Surveillance results demonstrated that kobuvirus and herpesvirus displayed obvious age distribution and PVs showed significant gender difference in YFPs. According to species demarcation criteria in individual genera in Papillomaviridae, two novel species (referred to as Omikronpapillomavirus 2 and 3) and four novel isolates of PV were identified in YFPs. Further evolutionary analysis suggested that NaPVs would occupy the mucosal niche and that virus-host codivergence mixed with duplications and host-switching events drives the evolution of cetacean PVs. Divergence times of PVs in YFP and other cetacean reflect the incipient speciation of YFPs. In summary, our findings revealed the potential viral entities, their prevalence, and their evolutionary history in YFPs, which raises an important issue regarding effects of viral infection on the fitness of YFPs. IMPORTANCE The Yangtze finless porpoise (YFP) is the only cetacean species in freshwater following the functional extinction of the baiji (Lipotes vexillifer). Health management, disease treatment, and other special measures are important for maintaining the existing YFP populations, especially in in situ and ex situ reserves. The discovery of potential viral entities and their prevalence in YFPs raises an important issue regarding the effects of viral infection on the fitness of YFPs and may contribute to the conservation of YFPs. The evolutionary history of papillomaviruses in YFP and other cetaceans reflects the phylogeny of their hosts and supports the status of incipient species, opening a window to investigate the evolutionary adaptation of cetaceans to freshwater as well as their phylogeny to remedy the deficiency of fossil evidence.

Reliability of caretakers' ratings of captive bottlenose dolphins (Tursiops truncatus) and finless porpoises' (Neophocaena asiaeorientalis sp.) behavioral style for welfare monitoring.

Qualitative ratings are increasingly used to assess animal welfare. We investigated the associations between daily caretakers' ratings of individuals' behavioral style, behavior during free time, participation during training sessions, and environmental variables in captive Yangtze finless porpoises, East-Asian finless porpoises, and bottlenose dolphins. Associations between caretakers' ratings and behavior were found, including social swimming, fast swimming, play, interaction with humans, and agonistic and socio-sexual interactions. Ratings were also associated with participation during training sessions and food intake. Social separation, transport to a new pool, high visitors' density, and holding of a sick conspecific were associated with ratings indicating a deteriorated human-animal relationship, a lower interest/fear of the environment, and/or a higher social cohesion depending on the group. Through principal component analyses, new variables that represented both caretakers' ratings and behavior and that could be used for an easier interpretation of the data for welfare monitoring within each group were extracted. Caretaker ratings and behavior should be used as a complement to previously validated indicators to detect changes in welfare state and understand the potential causes of these changes.

Seasonal Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis) movements in the Poyang Lake, China: Implications on flexible management for aquatic animals in fluctuating freshwater ecosystems.

Identifying seasonal high-use habitats and migration corridors is the basis for migratory species conservation. Previous studies have focused on Marine Protected Areas, while freshwater ecosystems, often accompanied by severe hydrological fluctuations, suggest new perspectives for flexible management. Poyang Lake is an essential habitat for the critically endangered Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis, YFP), supporting almost half of their natural population. However, studies on the movement patterns and habitat preferences of YFPs still lack there, preventing potential protected areas' effectiveness. In this study, we conducted 10 surveys for the YFP population in the main body and tributaries of Poyang Lake from October 2018 to August 2020. We used habitat modeling to analyze their seasonal habitat use and the environmental predictors. YFPs showed a seasonal movement pattern consistent with water level fluctuations. They entered the tributaries from the main lake body in low and medium water levels and returned to the main lake during high water level periods. The water depth was the most important environmental variable in each hydrological season that affects YFPs' habitat selection. The suitable water depth was about 4-8 m during the low water level, 6-12 m during the medium water level, and 7-20 m during the high water level. YFPs' 50% core habitats distribution showed noticeable seasonal changes. The proportion of their suitable habitats in the whole lake was relatively low, which was highest during high water level (16.89%), and the weakest during low water level (12.11%). Considering the seasonal movements of the YFPs in the lake, we recommend flexible management measures for their core habitats to alleviate human interference and restore their movement rhythm between the river and the lake, which shed light on protected area management for aquatic animals in such seasonal fluctuating habitats.

Blubber fatty acid compositions in different geographic populations of finless porpoise in Chinese waters: implications for thermal adaptation.

The stratification of blubber fatty acids (FAs) is a widely used strategy in marine mammals and can be influenced by many factors, including their diet, environmental temperature, and physiological status. There are 3 distinct finless porpoise species/subspecies in Chinese coastal and inland waters in a wide range from 20°N to 40°N. The biochemical stratification of the blubbers of finless porpoises in different regions may provide valuable information for understanding their environmental adaptations. The FA compositions of 4 geographic populations of finless porpoise (FP) collected from the Bohai Gulf, East China Sea, South China Sea, and Yangtze River were therefore analyzed and compared. The blubber FA compositions of finless porpoises were dominated by ΣMUFAs, followed by ΣSFAs and ΣPUFAs, and were generally consistent with those of other cetaceans. The blubber of finless porpoises was significantly stratified with increasing levels of ΣMUFAs and decreasing levels of ΣSFAs and ΣPUFAs from the inner to the outer layers. The 3 marine populations shared more similarities in their FA compositions and stratifications compared to the freshwater porpoises, particularly regarding the PUFAs in the inner layers, which might have been mainly influenced by the diet compositions of marine and freshwater porpoises. Contrary to what was expected, in the 3 marine populations, the SFA and MUFA levels showed opposite correlations (negative and positive, respectively) with habitat temperatures, possibly indicating an adaptation mechanism in finless porpoises characterized by a trade-off between the insulation and fluidity properties of the blubber through the adjustment of the compositions and gradients of MUFAs and SFAs across the blubber depth in response to environmental temperature/latitude changes.

The Click Production of Captive Yangtze Finless Porpoises (Neophocaena asiaeorientalis asiaorientalis) Is Influenced by Social and Environmental Factors.

Yangtze finless porpoises use high-frequency clicks to navigate, forage, and communicate. The way in which click production may vary depending on social or environmental context has never been investigated. A group of five captive Yangtze finless porpoises was monitored for one year, and 107 h of audio recordings was collected under different conditions. Using a MATLAB-generated interface, we extracted click density (i.e., number of clicks per minute) from these recordings and analyzed its variation depending on the context. As expected, click density increased as the number of animals present increased. The click density did not exhibit diurnal variations but did have seasonal variations, with click density being highest in summer and fall. Yangtze finless porpoises produced more clicks when socially separated than when not (136% more), during training/feeding sessions than outside of such sessions (312% more), when enrichment was provided (265% more on average), and when noisy events occurred rather than when no unusual event occurred (22% more). The click density decreased when many visitors were present in the facility (up to 35% less). These results show that Yangtze finless porpoises modulate their click production depending on the context and suggest that their echolocation activity and their emotional state may be linked to these changes. Such context-dependent variations also indicate the potential usefulness of monitoring acoustical activity as part of a welfare assessment tool in this species. Additionally, the click density variation found in captivity could be useful for understanding click rate variations of wild populations that are hardly visible.

Sexual maturity, seasonal estrus, and gestation in female Indo-Pacific bottlenose dolphins Tursiops aduncus inferred from serum reproductive hormones.

Reproductive hormones in serum concentrations of progesterone, estradiol, and testosterone in female Indo-Pacific bottlenose dolphins (Tursiops aduncus, n = 12) housed in Ocean Park Hong Kong were investigated in the present study. Results showed that, onset of puberty of captive Indo-Pacific bottlenose dolphins was at 5 years while sexual maturity was at 6. Average serum progesterone concentrations in non-pregnant sexually mature individuals was 0.33 (0.25-0.97) ng/mL (interquartile), significantly higher than in immature ones 0.26 (0.25-0.38) ng/mL. This study found significant difference in serum estradiol concentrations between individuals at the onset of puberty (9.5 ± 1.7 pg/mL, ±SD) and not (below detection limit 9 pg/mL). A slightly seasonal breeding pattern, with progesterone values tend to be higher from February to October (0.38 [0.25-1.07] ng/mL) was inferred. During pregnancy, serum progesterone concentrations range from 10.54 ± 8.74 ng/mL (indexed month post-conception [IMPC] 0) to 25.49 ± 12.06 ng/mL (IMPC 2), and display a bimodal pattern with 2 peaks in early- (25.49 ± 12.06 ng/mL, IMPC 2) and late-pregnancy (21.71 ± 10.25 ng/mL, IMPC 12), respectively. Serum estradiol concentrations can seldom be detected in early-pregnancy and increase constantly in mid- (9.45 ± 1.83 pg/mL) and late-pregnancy (11.88 ± 3.81 pg/mL), with a spike (15.45 ± 6.78 pg/mL) 1 month prior to delivery. Serum testosterone concentrations elevate significantly in IMPC 7 (0.36 ± 0.10 ng/mL) compared to other months (0.16 ± 0.10 ng/mL) of the year. The present study provides normal concentration profiles for some reproductive hormones in female Indo-Pacific bottlenose dolphins and can contribute to the breeding monitoring of this species. Also, our study would shed further light on the reproductive physiology of small cetaceans.