中华白海豚雄性生殖系统组织学初步研究

采用组织学和形态学方法研究了中华白海豚(Sousa chinensis)的雄性生殖系统。以睾丸和附睾中是否存在精母细胞及精子细胞等作为判断雄性中华白海豚性成熟的标准,通过比较不同个体睾丸和附睾的组织结构特征,发现成熟个体的生精小管和附睾的组织学结构与未成熟个体间存在显著差异。通过测量样本睾丸的2个形态参数:生精小管直径和生精小管的相对面积,得到性成熟个体的生精小管直径为(118.3±12.8)μm,生精小管相对面积为0.52;未成熟个体的生精小管直径(47.4±3.5)～(60.3±6.0)μm,生精小管相对面积为0.27～0.40。     

Oxidation-reduction potentials and spectral properties of some cytochromes from Thiobacillus versutus (A2)

Antibodies raised against rat hepatic epoxide hydrolase (EC 3.3.2.3) and glutathione S-transferases (EC 2.5.1.18) B, C and E were used to determine the presence and localizations of these epoxide-metabolizing enzymes in testes of sexually immature and mature Wistar and Holtzman rats. Unlabeled antibody peroxidase-antiperoxidase staining for each enzyme was readily detected in rat testes at the light microscopic level. Although significant strain-related differences were not apparent, staining intensity for certain enzymes differed markedly between Leydig cells and seminiferous tubules. Leydig cells of immature and mature rats were stained much more intensely for epoxide hydrolase and glutathione S-transferases B and E than were seminiferous tubules, whereas Sertoli cells, spermatogonia, spermatocytes and spermatids, as well as Leydig cells, were stained intensely by the anti-glutathione S-transferase C. Age-related differences in staining for glutathione S-transferase B were not obvious, while the anti-glutathione S-transferase C stained seminiferous tubules more intensely in immature rats, and antibodies to epoxide hydrolase and glutathione S-transferases C and E stained Leydig cells much more intensely in mature rats. These observations thus demonstrate that testes of both sexually immature and mature rats contain epoxide hydrolase and glutathione S-transferases. Except for glutathione S-transferase C in immature rats, Leydig cells appear to contain much higher levels of enzymes than do seminiferous tubules. During sexual maturation, the testicular level of glutathione S-transferase B appears to remain constant, while levels of epoxide hydrolase and glutathione S-transferases C and E increase within Leydig cells and the level of glutathione S-transferase C decreases within seminiferous tubules.     

Strandings provide insight into social group structure of Atlantic white-sided dolphins

Atlantic white-sided dolphins (Lagenorhynchus acutus) are highly social odontocetes with a poorly understood tendency to mass strand. With limited capacity to study social ecology in the open ocean, mass strandings provide an opportunity to improve our understanding of group structure. Our study of 32 mass stranding events that occurred on Cape Cod, Massachusetts, between 1999 and 2009 identifies aspects of social ecology that vary across the year. A greater number of mass stranding events occurred outside of the breeding season and there was evidence of age-structuring during the breeding season. We find generally low average intragroup pairwise relatedness assessed across eight microsatellite loci in a subset of 16 mass stranding events. Mass stranded groups do not show higher than expected relatedness when compared to a baseline estimate derived from single-stranded individuals. Overall, our integration of genetic estimates of relatedness with data on sex and maturity-class from stranded specimens suggests that Atlantic white-sided dolphins fall near the more fluid end of the continuum from short-term, highly fluid social associations to long-term, stable groups represented among the odontocetes. Despite their tendency to mass strand, stable, kin-based associations are not a defining feature of social group structure in this species.     

Localization of epoxide-metablozing enzymes in rat testis

Antibodies raised against rat hepatic epoxide hydrolase (EC 3.3.2.3) and glutathione S-transferases (EC 2.5.1.18) B, C and E were used to determine the presence and localizations of these epoxide-metabolizing enzymes in testes of sexually immature and mature Wistar and Holtzman rats. Unlabeled antibody peroxidase-antiperoxidase staining for each enzyme was readily detected in rat testes at the light microscopic level. Although significant strain-related differences were not apparent, staining intensity for certain enzymes differed markedly between Leydig cells and seminiferous tubules. Leydig cells of immature and mature rats were stained much intensely for epoxide hydrolase and glutathione S-transferase B and E than were seminiferous tubules, whereas Sertoli cells, spermatogonia, spermatocytes and spermatids, as well as Leydig cells, were stained intensely by the anti-glutathione S-transferase C. Age-related differences in staining for glutathione S-transferase B were not obvious, while the anti-glutathione S-transferase C stained seminiferous tubules more intensely in immature rats, and antibodies to expoxide hydrolase and glutathione S-transferases C and E stained Leydig cells much more intensely in mature rats. These observations thus demonstrate that testes of both sexually immature and mature rats contain epoxide hydrolase and glutathione S-transferases. Except for glutathione S-transferase C in immature rats, Leydig cells appear to contain much higher levels of enzymes than do seminiferous tubules. During sexual maturation, the testicular level of glutathione S-transferase B appears to remain constant, while levels of epoxide hydrolase and glutathione S-transferases C and E increase within Leydig cells and the level of glutathione S-transferase C decreases within seminiferous tubules.     

Microbiological characterization of three Atlantic whiteside dolphins (Lagenorhynchus acutus) from stranding through captivity with subsequent rehabilitation and release of one animal

Three live Atlantic whiteside dolphins (Lagenorhynchus acutus) were found beach-stranded on Cape Cod in Massachusetts and were shipped to Mystic Marinelife Aquarium in Connecticut and held in captivity. Swab samples were collected routinely from the anus and blowhole of live animals and from internal organs at necropsy for microbial culture. Despite antibiotic therapy, one animal died on day 9 of captivity, and one died after 110 days. The survivor was released after 126 days of captivity. Klebsiella pneumoniae septicemia and staphylococcal pneumonia were likely responsible for the two deaths. A total of 26 different organisms (or groups of organisms) were recovered from the three dolphins, including several (e.g., Pseudomonas spp, Streptococcus faecalis Candida albicans, Proteus-Morganella spp, Vibrio spp) that, in addition to the above organisms, could be potentially pathogenic for debilitated animals. Newly rescued marine mammals should be segregated from healthy animals if possible.     

Clinical assessment and postrelease monitoring of 11 mass stranded dolphins on Cape Cod,Massachusetts

The health, postrelease movements, and behavior of mass stranded Atlantic white‐sided dolphins (Lagenorhynchus acutus) and short‐beaked common dolphins (Delphinus delphis) from Cape Cod, Massachusetts, were evaluated. Health was assessed through physical examination and blood analysis. Eleven dolphins (eight white‐sided dolphins and three common dolphins) were relocated, outfitted with satellite transmitters, and released during seven mass stranding events. Five transmitters recorded only location, and six also included a time‐depth recorder. Transmission duration ranged from 8 h to 218 d, with a mean of 117 d (median = 118 d, SD = 82 d), after release. All dolphins demonstrated extensive movement throughout the Gulf of Maine. The distribution of tagged dolphins was considered normal based on comparisons with published data for these species. Excluding the dolphin that transmitted for only 8 h, mean minimum speeds for individual dolphins ranged from 3.4 to 6.6 km/h; overall mean for all dolphins was 5.4 km/h (SD = 0.9 km/h). The five dolphins with time‐depth recorders had mean dive depths of 8.6–40.3 m and mean dive durations of 46–296 s. Hematologic and biochemical data revealed only minor abnormalities. Data suggest that at least 10 of the 11 dolphins were likely successfully reintroduced into the wild.     

Features of impaired seminiferous tubule differentiation are associated with germ cell neoplasia in adult men surgically treated in childhood because of cryptorchidism

Seminiferous tubule differentiation was related to the occurrence of germ cell neoplasia in 38 men, aged 17-47, treated surgically in childhood for cryptorchidism. Tissues from 46 testes obtained from biopsies taken as a neoplastic preventive procedure or whole testes removed because of GCT were evaluated quantitatively. Paraffin sections were treated with antibodies against placental like alkaline phosphatase (PLAP), a marker of germ cell neoplasia, and cytokeratin 18 (CK-18), a marker of immature Sertoli cells. Quality of spermatogenesis and number Leydig cells were assessed with a score count. Seminiferous tubules diameter, thickness of basal membrane and size of intertubular spaces were measured with image analysis software. In 17.4% of testes spermatogenesis was normal (9.9 points) (N) and neoplasia was not found there. In the other 38 specimens (83%) spermatogenesis was abnormal (A). When spermatogenesis was arrested or when germ cells were absent (3.7+/-1.8 points), neoplastic lesions were found in 13.1% of the specimens. In A group 5.1+/-7.1% of tubules contained immature Sertoli cells, while in N they were not found. Tubular diameter was significantly lower in A (161.5+/-31.8 microm) than in N (184.6+/-24.3 microm) and the percentage of seminiferous tubules with the thickening of tubular basal membrane was also greater in A. Intertubular spaces were significantly larger in A (49.9+/-18.6%) in comparison to N group (32.6+/-12.5%). Mean number of Leydig cells was similar in both groups. To conclude, in most of the formerly cryptorchid testes, despite surgical treatment, impaired seminiferous tubules differentiation is predominant. Germ cell neoplasia is present in testes with retarded seminiferous tubules differentiation. Retardation of seminiferous tubule differentiation consists of inhibited spermatogenesis, presence of tubules with immature Sertoli cells, decreased tubular diameter, increased thickness of basal membrane and enlarged intertubular spaces. Examination of testicular biopsy with respect to the state of seminiferous tubule differentiation may be helpful to predict the appearance of germ cell neoplasia in adult men with cryptorchidism in anamnesis. Orchiopexy of cryptorchid testes may not prevent the occurrence of features of testicular dysgenesis and the associated germ cell neoplasia.     

A genomewide catalogue of single nucleotide polymorphisms in white‐beaked and Atlantic white‐sided dolphins

The field of population genetics is rapidly moving into population genomics as the quantity of data generated by high‐throughput sequencing platforms increases. In this study, we used restriction‐site‐associated DNA sequencing (RADSeq) to recover genomewide genotypes from 70 white‐beaked (Lagenorhynchus albirostris) and 43 Atlantic white‐sided dolphins (L. acutus) gathered throughout their north‐east Atlantic distribution range. Both species are at a high risk of being negatively affected by climate change. Here, we provide a resource of 38 240 RAD‐tags and 52 981 nuclear SNPs shared between both species. We have estimated overall higher levels of nucleotide diversity in white‐sided (π = 0.0492 ± 0.0006%) than in white‐beaked dolphins (π = 0.0300 ± 0.0004%). White‐sided dolphins sampled in the Faroe Islands, belonging to two pods (N = 7 and N = 11), showed similar levels of diversity (π = 0.0317 ± 0.0007% and 0.0267 ± 0.0006%, respectively) compared to unrelated individuals of the same species sampled elsewhere (e.g. π = 0.0285 ± 0.0007% for 11 Scottish individuals). No evidence of higher levels of kinship within pods can be derived from our analyses. When identifying the most likely number of genetic clusters among our sample set, we obtained an estimate of two to four clusters, corresponding to both species and possibly, two further clusters within each species. A higher diversity and lower population structuring was encountered in white‐sided dolphins from the north‐east Atlantic, in line with their preference for pelagic waters, as opposed to white‐beaked dolphins that have a more patchy distribution, mainly across continental shelves.     

Cadmium-containing granules in kidney tissue of the Atlantic white-sided dolphin (Lagenorhyncusacutus) off the Faroe Islands

Top predators from the northern sub-polar and polar areas exhibit high cadmium concentrations in their tissues. In the aim to reveal possible adverse effects, samples of five Atlantic white-sided dolphins Lagenorhyncusacutus have been collected on the occasion of the drive fishery in the Faroe Islands, for ultrastructural investigations and energy dispersive X-ray microanalyses. Cadmium concentrations were less than the limit of detection in both immature individuals and ranged from 22.7 to 31.1 μg g^?1 wet weight in the mature individuals. Two individuals with the highest cadmium concentrations exhibited electron dense mineral concretions in the basal membranes of the proximal tubules. They are spherocrystals made up of numerous strata mineral deposit of calcium and phosphorus together with cadmium. Cadmium has been detected with a molar ratio of Ca:Cd of 10:1 in the middle of these concretions. To our knowledge, this is the first report of such granules in a wild vertebrate. The role of these granules in the detoxification of the metal and the possible pathological effects are considered.     

Impact of fine needle aspiration (FNA) and of the number of punctures on the feline testis: clinical, gross anatomy and histological assessment

The safety of testicular fine needle aspiration (FNA) has been proven in dogs but has not been fully established in men, while studies in rats have given contradictory results. Furthermore, the extent of damage inflicted by multiple punctures is unknown. The aim of this study was to determine the impact of FNA and of the number of punctures on the feline testis with clinical, gross anatomy and histological examinations. Twenty-seven sexually mature healthy laboratory Domestic Shorthair cats were randomly assigned to two groups: 5 cats in which no FNA was performed (control group), and 22 cats which had their left and right testis punctured with a 26 ga needle towards 3 and 8 directions, respectively (experimental group). Two cats at a time were orchiectomized 5 or 30 min, 1, 2, 4, 7 or 14 days or 1, 2, 3 or 4 mo post-aspiration. The cats of the control group were also orchiectomized. During the first week post-aspiration clinical examination revealed vaginal cavity hematoma (8/44 testes), while the histological findings were focal hemorrhagic areas (20/24 testes), erythrocytes inside the seminiferous tubules' lumen (9/24 testes), and germinal cell degeneration in <1.94% of the seminiferous tubules (15/24 testes). After the first week the histological findings were germinal cell degeneration in <2.14% of the seminiferous tubules (19/20 testes) and enlargement of the lumen of <5.16% of the seminiferous tubules (7/20 testes). The germinal epithelium and interstitium had an overall normal appearance. No significant differences were observed between the left and right testis. The results of the study indicate that testicular FNA should be considered a safe procedure in the cat when up to 8 punctures are performed.     

Reproduction of the blacktip sawtail catshark,Galeus sauteri,in the waters off northeastern Taiwan

The reproductive biology of blacktip sawtail catsharks,Galeus sauteri, in northeastern Taiwan waters was investigated. Male catsharks possessed paired testes producing spermatozoa, which were then stored in the epididymides all year round. No spermatophores were observed in the lower ductus deferens. Only the right ovary was functional in females, oogonia being formed in the ovarian cortex and then developing into mature ova. Ova exceeding 17 mm in diameter were ovulated. Pregnant individuals contained one eggcase only, in each side of the uterus. No hatched embryos were observed in such eggcases. The size at which 50% of blacktip sawtail catshark specimens were mature was 410–420 mm and 350–360 mm for females and males, respectively. The species does not have a well-defined reproductive season.     

Reproductive condition of Akodon montensis Thomas and Bolomys lasiurus (Lund) (Rodentia, Muridae) based on histological and histometric analyses of testes and external characteristics of gonads

The reproductive activity of two species of wild rodents was studied in the Serra do Rola‐Moça State Park (PERM), in the state of Minas Gerais, Brazil. From October 1998 to October 1999 specimens of Akodon montensis (Thomas 1913) were captured in an area of seasonal semi‐deciduous forest, while specimens of Bolomys lasiurus (Lund 1840) were captured in a cerrado area (a savannah‐like grassland). The determination of the reproductive condition was based on histological and histometric analyses of testes and external characteristics of gonads. The volume of the testes, diameter of the seminiferous tubules and depth of the seminiferous epithelium were measured. Akodon montensis showed a seasonal annual reproductive cycle divided into four stages (recrudescence, activity or proliferation, regression and rest) with the reproductive activity mainly occurring during the rainy season. In contrast, B. lasiurus presented reproductive activity during almost the whole year. For both species the volume of the testes was an effective indicator of their reproductive condition. We suggest that the histological and histometric analysis of the gonads of males, together with the observations of external characteristics of gonads of males and females, may assist in identifying the reproductive condition of these animals.     

Histochemical detection of steroid synthesizing cells in the testes of goldfish,Carassius auratus,during the annual reproductive cycle

We investigated the sites of Δ⁵-3β-hydroxysteroid dehydrogenase (3 β-HSD) and glucose-6-phosphate dehydrogenase (G-6-PD) synthesis in the testes of goldfish, Carassius auratus, during the annual reproductive cycle. The histochemistry of fish gonads has been investigated previously in many species other than goldfish. The reproductive cycle of goldfish, is divided into five stages and the steroid synthesizing cells of the testes were studied during these stages, using histochemical techniques. We found that interstitial cells and seminiferous tubules are the main steroid synthesizing sites in testes of goldfish, and that enzyme activity was more intense in the interstitial cells than in the seminiferous tubules. During the pre-spawning months, 3 β-HSD and G-6-PD activities were weak compared to the spawning months.     

Age and growth in Peale's dolphin (Lagenorhynchus australis) in subantarctic waters off southern South America

The Peale's dolphin, Lagenorhynchus australis, is a mainly coastal and one of the most often seen species off southernmost South America from 33°S off Chile and 38°S off Argentina to south of Cape Horn. Although a common species, its IUCN status is “Data Defficient”, so any information on this species is relevant. Age, growth and physical maturity were examined in skeletons of 57 specimens of this species, mostly from the coasts of Tierra del Fuego, Argentina; this represents most of the known specimens of this species available in museum and private collections. Most specimens were by-catch in shore-set gillnets or had been harpooned for crab bait. Age was determined for 23 specimens, all from Tierra del Fuego. The maximum age was 13 years for a 199 cm female. The largest female in this survey was 210 cm, the largest male 205 cm in total length (12 years), but this probably does not represent the maximum length for this species. Only two animals, of 12 and 13 years, were physically mature. Animals reach asymptotic length (188 cm) with less than the 30% of the vertebral epiphyses fused. The animals in this study were mainly subadults, as has been found for other southern by-caught dolphins. Growth was studied for total length using 18 osteological characters with the Gompertz model. We propose that zygomatic width can be used to estimate total length for incomplete beach-cast specimens.     

Taenia crassiceps: Infections of male mice lead to severe disruption of seminiferous tubule cells and increased apoptosis

This research was carried out to study the effects of infection with Taenia crassiceps cysticerci on the seminiferous epithelium histoarchitecture in the testes of male mice. Our results showed a severe disruption of the histoarchitecture of the testis epithelium in infected mice. In these animals, a significant infiltration of macrophages within seminiferous tubules was observed (P < 0.001). Generalized apoptosis of germ cells within the seminiferous tubules was observed, as assessed by TUNEL assay and apoptotic nuclei were quantified. The total number of fluorescent objects (DNA) (including clusters, singles, and objects in clusters) was significantly higher in the infected cells than in the control group (P = 0.0286). Observation of the interstitial tissue showed disorder and deterioration of many Leydig cells of infected mice, as well as intense vacuolization and destruction of their inter-cellular junctions. Several ultrastructural abnormalities were observed through electron microscopy as well. The observed pathology could lead to a state of infertility.     

Biphonal calls in Atlantic spotted dolphins (Stenella frontalis): bitonal and burst-pulse whistles

Biphonation, the simultaneous production of two sounds by a single animal, is found in the vocalizations of a range of mammalian species. Its prevalence suggests it plays an important role in acoustic communication. Concurrent vocal and behavioural recordings were made of Atlantic spotted dolphins (Stenella frontalis) off Bimini, The Bahamas. The occurrence of two types of biphonal signals is reported: burst-pulse whistles with combined tonal and burst-pulse elements, and bitonal whistles. Biphonal whistles are rarely described in reports of dolphin acoustic repertoires, but were common in these dolphins: of all whistles analysed (n = 1211), 26.84% were burst-pulse whistles and 4.71% were bitonal whistles. A subset of whistles (n = 397) were attributed to dolphins of specific age classes, and used to compare prevalence of biphonation across age. Biphonation occurred in 61.54% of sexually mature and 48.32% of sexually immature dolphins’ whistles. Sexually immature dolphins emitted more burst-pulse whistles than older dolphins: 44.13% of sexually immature dolphins’ whistles were burst-pulse whistles, while 15.38% of adult whistles were burst-pulse whistles. Bitonal whistle production was more prevalent in sexually mature dolphins: 41.03% of adult whistles were bitonal, while only 4.19% of sexually immature dolphins’ whistles were bitonal. The prevalence of a biphonal component in specific repeated, stereotyped whistle contours suggests that these acoustic features could be important components of contact calls, or signature whistles. The biphonal components of spotted dolphin whistles may serve to convey additional information as to identity, age or other factors to conspecifics.     

Temporal germ cell development strategy during spermatogenesis within the testis of the Ground Skink, Scincella lateralis (Sauria: Scincidae)

Ground Skink (Scincella lateralis) testes were examined histologically to determine the testicular organization and germ cell development strategy employed during spermatogenesis. Testicular tissues were collected from 19 ground skinks from Aiken County, South Carolina during the months of March-June, August, and October. The testes consisted of seminiferous tubules lined with germinal epithelia in which germ cells matured in close association with Sertoli cells. As germ cells matured, they migrated away from the basal lamina of the epithelia towards the lumina of the seminiferous tubules. The testes were spermatogenically active during the months of March, April, May, June, and October (largest seminiferous tubule diameters and epithelial heights), but entered a quiescent period in August (smallest seminiferous tubule diameter and epithelial height) where only spermatogonia type A and B and early spermatocytes were present in low numbers within the seminiferous epithelium. Although the testicular organization was similar to other amniotes, a temporal germ cell development strategy was employed during spermatogenesis within Ground Skinks, similar to that of anamniotes. Thus, this skink's germ cell development strategy, which also has been recently reported in all other major reptilian clades, may represent an evolutionary intermediate in terms of testicular organization between anamniotes and birds and mammals.     

Seasonal spermatogenic cycle and morphology of germ cells in the viviparous lizard Mabuya brachypoda (Squamata,Scincidae)

We describe seasonal variations of the histology of the seminiferous tubules and efferent ducts of the tropical, viviparous skink, Mabuya brachypoda, throughout the year. The specimens were collected monthly, in Nacajuca, Tabasco state, Mexico. The results revealed strong annual variations in testicular volume, stages of the germ cells, and diameter and height of the epithelia of seminiferous tubules and efferent ducts. Recrudescence was detected from November to December, when initial mitotic activity of spermatogonia in the seminiferous tubules were observed, coinciding with the decrease of temperature, photoperiod and rainy season. From January to February, early spermatogenesis continued and early primary and secondary spermatocytes were developing within the seminiferous epithelium. From March through April, numerous spermatids in metamorphosis were observed. Spermiogenesis was completed from May through July, which coincided with an increase in temperature, photoperiod, and rainfall. Regression occurred from August through September when testicular volume and spermatogenic activity decreased. During this time, the seminiferous epithelium decreased in thickness, and germ cell recruitment ceased, only Sertoli cells and spermatogonia were present in the epithelium. Throughout testicular regression spermatocytes and spermatids disappeared and the presence of cellular debris, and scattered spermatozoa were observed in the lumen. The regressed testes presented the total suspension of spermatogenesis. During October, the seminiferous tubules contained only spermatogonia and Sertoli cells, and the size of the lumen was reduced, giving the appearance that it was occluded. In concert with testis development, the efferent ducts were packed with spermatozoa from May through August. The epididymis was devoid of spermatozoa by September. M. brachypoda exhibited a prenuptial pattern, in which spermatogenesis preceded the mating season. The seasonal cycle variations of spermatogenesis in M. brachypoda are the result of a single extended spermiation event, which is characteristic of reptilian species. J. Morphol. © 2012 Wiley Periodicals, Inc.