Identification of sperm subpopulations with defined motility characteristics in ejaculates from Ile de France rams

The aims of this study were to identify different motile sperm subpopulations in fresh ejaculates from six Ile de France rams, by using a computer-assisted sperm motility analysis (CASA) system, and to evaluate the effects of individual ram and season on population distribution. Overall sperm motility and individual kinematic parameters of motile spermatozoa were evaluated for 125,312 spermatozoa, defined by curvilinear velocity (VCL), linear velocity (VSL), average path velocity (VAP), linearity coefficient (LIN), straightness coefficient (STR), wobble coefficient (WOB), mean amplitude of lateral head displacement (ALH) and frequency of head displacement (BCF). A multivariate cluster analysis was carried out to classify these spermatozoa into a reduced number of subpopulations according to their movement patterns. The statistical analysis clustered the whole motile sperm population into five separate groups: subpopulation 1, constituted by rapid, progressive and non sinuous spermatozoa (VCL=126.41 μm/s, STR=92.87% and LIN=86.47%); subpopulation 2, characterized by progressive spermatozoa with moderate velocity (VCL=74.74 μm/s and STR=84.03%); subpopulation 3, represented by rapid, progressive and sinuous spermatozoa (VCL=130.45 μm/s, STR=76.02% and LIN=47.68%); subpopulation 4 represents rapid nonprogressive spermatozoa (VCL=128.69 μm/s and STR=44.09%); subpopulation 5 includes poorly motile, nonprogressive spermatozoa with a very irregular trajectory (VCL=36.81 μm/s and STR=47.04%). Our results show the existence of five subpopulations of motile spermatozoa in ram ejaculates. The frequency distribution of spermatozoa within subpopulations was quite similar for the six rams, and the five subpopulations turned out to be very stable along seasons.     

Aspartate aminotransferase in ejaculates of Manchego and Merino rams after minimal and maximal sperm damage

Semen from Manchega and Merina breed rams was subjected to minimal and maximal sperm cell damage treatments. Seminal plasma samples were then analyzed for aspartate aminotransferase (AAT) activity. There were significant (P < 0.01) increases in enzymatic activity after maximal damage treatment in both breeds. Mean values of AAT activity were different between months (P < 0.01) and individual rams (P < 0.01). Breed significantly (P < 0.01) affected AAT activity after minimal and maximal sperm damage. There was an interaction (P < 0.01) between the 2 factors of ram and month in both breeds, but there was no interaction between breed and treatment.     

Treponematosis in regional and chronological perspective from central Gulf Coast Florida

Examination of five central Gulf Coast Florida archaeological skeletal series (n = 547) from the late prehistoric and early historic periods reveals the presence of pathological lesions characteristic of treponemal infection. These skeletal remains of native American populations represent a crucial time regarding hypotheses about the exchange of syphilis between the Old and New World or the mutation of one treponemal infection into another. Comparison of the lesions with those observed in modern studies of treponemal infection does not support an interpretation of veneral syphilis. The data from this study and from other archaeological skeletal series indicate the presence of a treponemal disease prior to European contact. The skeletal elements from Florida do not suggest that the disease changed dramatically following contact with Europeans. It is possible that cases of nonspecific postcranial pathology can be explained by the presence of this disease in the Florida populations. This study indicates that a treponemal disease was endemic throughout the region by at least A. D. 1000. © 1993 Wiley-Liss, Inc.     

Cooling and freezing of sperm from captive,free-living and endangered squirrel monkey species

Germoplasm banking is an important tool for the preservation of genetic material from Neotropical primates in captivity, and from free living species, especially the endangered ones like Saimiri vanzolinii (Black-headed squirrel monkey), a primate with a low incidence area (870 km² of floodplains) in the southern part of the Mamirauá Sustainable Development Reserve, Brazil. Therefore, in the present study we aimed to develop a sperm cryopreservation protocol comparing sperm cooling in presence (T1) and absence (T2) of egg yolk, and to test freezing protocols to preserve semen from captive (Saimiri collinsi), and free-living (Saimiri vanzolinii, Saimiri cassiquiarensis and Saimiri macrodon) New World primates. Cooling preserved sperm of S. collinsi in all evaluated microscopic parameters, except for sperm motility. No differences were observed among the treatments, indicating that semen of this species can be cooled without egg yolk. Freezing did not affect sperm quality of S. collinsi, except plasma membrane integrity that was negatively affected. Generally, a good maintenance rate was observed between cooling and thawing of semen for the four species, showing the positive translational application of protocols from S. collinsi to the free-living species. Developed freezing protocol proved to be useful for sperm cryopreservation of S. collinsi and in field conditions.     

Effects of freezing/thawing on motile sperm subpopulations of boar and donkey ejaculates

The main aim of this study is to assess the influence of freeze/thawing on motile sperm subpopulations in ejaculates from two phylogenetically different mammalian species, boar and donkey. Our results indicate that, whereas boar and donkey sperm respond very differently in their mean motion characteristics to freezing/thawing, this process did not change the existence of a 4-subpopulations structure in the ejaculates in either species when these subpopulations were defined by taking values of curvilinear velocity (VCL) as reference. Moreover, the freezing/thawing-linked changes in mean sperm-motion characteristics in both boar and donkey semen were especially due to changes in the proportion among each concrete subpopulation. In this way, the freezing/thawing-induced mean increase in motion characteristics observed in boar sperm was a result of the decrease in the percentage of sperm in Subpopulation 1 (from 53.9%+/-4.7% to 31.2%+/-3.9% after thawing) and a concomitant increase of sperm from Subpopulations 3 (from 13.3%+/-2.5% to 32.6%+/-3.9% after thawing) and 4 (from 3.4%+/-0.9% to 8.0%+/-1.1% after thawing). On the contrary, changes in mean motility of frozen/thawed donkey sperm were linked to an increase in the percentage of sperm in Subpopulation 1 (from 31.5%+/-4.3% to 58.8%+/-4.9% after thawing) and a concomitant decrease of sperm from Subpopulations 3 (from 32.4%+/-3.2% to 6.6%+/-1.8% after thawing) and 4 (from 12.2%+/-2.5% to 7.3%+/-1.9% after thawing). In conclusion, our results seem to indicate that motility changes induced by the freezing/thawing protocol are linked to concomitant changes in both the specific parameters and, more importantly, to the specific percentage of each of the motile sperm subpopulations. These changes did not affect the overall proportion of motile sperm present in both boar and donkey, which is conserved despite the detrimental effect caused by freezing/thawing in both species. Finally, the presence of some kind of motile sperm subpopulations structure has been described in mammalian species with a very great phylogenetic distance, thus suggesting that this structure could play some role in the maintenance of the overall function of mammalian ejaculates.     

Functional parameters of chimpanzee (Pan troglodytes) sperm from ejaculates collected by rectal probe electrostimulation and by artificial vagina

This study compares functional parameters of sperm from ejaculates collected from 15 adult male chimpanzees using rectal probe electrostimulation (RPE) and from 10 adult male chimpanzees trained to use an artificial vagina (AV). Computer assisted motion analysis (CAMA) showed no significant differences in mean values for straight line velocity (VSL), linearity (LIN), curvilinear velocity (VCL), and lateral head movement (ALH) of sperm from ejaculates collected by RPE and by AV. There was, however, a significant difference (P < 0.01) in the population distribution for VSL and LIN, which indicates that sperm swim in a more convoluted manner in ejaculates collected by RPE than in ejaculates collected by AV. In the hamster zona-free ovum penetration assay (SPA), there were no significant differences in the percentages of hamster oocytes penetrated by sperm or in the number of sperm which penetrated each oocyte after 4 or 24 h incubation using sperm from ejaculates collected by RPE and by AV. Therefore, the lack of success using sperm from ejaculates collected by RPE to initiate pregnancy in the chimpanzee does not appear to result from abnormalities in sperm fertilizing capacity as measured in SPA. © 1996 Wiley-Liss, Inc.     

北部湾徐闻海域红树内生细菌物种多样性及其杀线虫活性研究

为发掘红树植物内生细菌资源和寻找新型微生物杀虫剂,该研究从北部湾徐闻海域采集7种红树植物共16份样品,设计10种分离培养基,使用稀释涂布法分析红树植物内生细菌的分布特征,通过16S rRNA分子生物学方法对内生细菌进行多样性分析,并利用秀丽隐杆线虫模型,通过杀线虫活性实验测试内生细菌乙酸乙酯提取物的杀线虫活性。结果表明:(1)从16份红树植物各组织器官中获得33株内生细菌,分布于19个科23个属。其中,芽孢杆菌属(Bacillus)为优势菌属,并发现了10株潜在的新种或新属。(2)筛选到具有显著杀线虫活性的菌株IMDGX 4725和IMDGX 4744,半数致死浓度(LC₅₀)分别为61.58、100.89mg·mL^-1。研究结果证实了徐闻海域红树具有多样性丰富的内生细菌,同时部分细菌具有较强的杀线虫活性,具有发现新型微生物杀虫剂的潜力。     

Paleoecology of epizoans and borings on some Upper Cretaceous chalk oysters from the Gulf Coast

Analysis was made of a hard substrate fauna found on right valve interiors and exteriors of the epifaunal reclining oyster Pycnodonte mutabilis from the Maastrichtian (Navarroan) Saratoga Formation (southwestern Arkansas). Comparison of boring and encrustation patterns on both sides of valves indicates that a major portion of colonization on valve exteriors occurred while host oysters were alive. Paleoautecologic information derived from such valve exterior patterns includes evidence of rheotropic orientation by encrusting juvenile P. mutabilis and preferential location of Trypanites sp. borings in surficial shell grooves. Valve exteriors supported a hard substrate paleocommunity which had the following non-interactive progressive colonization sequence: (1) Trypanites sp. and P. mutabilis juveniles; (2) Entobia sp., serpulid worm tubes, and Bullopora sp.; and (3) cheilostome bryozoans. This sequence could have been caused by low seasonality and ranked success of colonizing encrusters and borers. Colonization of valve interiors generally differed from exteriors only in that many interiors were first colonized by the clionid sponge that created Entobia sp., which had already occupied the exterior, and which quickly bored through the valve to occupy the interior upon the host's death. □ Trace fossils, epizoans, borings, Gryphaeidae, palaeoecology, communities, colonization sequence, Late Cretaceous, Maastrichtian, Navarroan, Arkansas.     

Cryopreservation of semen from endangered pheasants: the first step towards a cryobank for endangered avian species

In order to improve the genetic management of bird species within the European Endangered Programs (EEP), a research project on artificial insemination and cryopreservation of Galliformes semen has been developed. The aim of the program is to create a sperm cryobank for threatened bird species. During this study, semen was collected from 17 pheasant species and specific characteristics of ejaculates were analyzed (volume, sperm concentration, motility, pH). Artificial insemination with fresh semen was performed in nine species and with frozen semen in eight species. Inseminations with frozen and thawed semen were made in 17 species. Viability of fresh and frozen semen was assessed in vitro using double stains, eosin and nigrosin. The effect of pH (7-8.5) on viability of fresh and frozen/thawed spermatozoa was also studied. Chicks hatched in eight and three species after insemination with fresh and frozen/thawed semen, respectively. Species varied widely in semen viability: 1-30% of spermatozoa survived freezing and thawing. There was a negative correlation between the viability of frozen spermatozoa and semen pH. In our experimental conditions, the pH of diluents had no effect on semen viability. However, semen with the highest pH had the lowest quality after freezing and thawing. These experiments demonstrated the feasibility of using a very simple and inexpensive method to achieve artificial insemination and cryopreservation of semen in endangered pheasant species.     

Fatty acid contents and profiles of 16 macroalgae collected from the Irish Coast at two seasons

Due to the established health benefits of omega-3 long-chain polyunsaturated fatty acids (LC-PUFA), there is a globally increasing demand for alternative natural resources with appropriate fatty acid profiles. To assess the suitability of macroalgae as a source, 16 species (nine Phaeophyceae, five Rhodophyta and two Chlorophyta) were collected at two seasons (June and November) from the Irish west Coast, and total fatty acid contents and specific profiles were determined. Total fatty acid contents, expressed per percentage of dry weight, ranged from 6.4 %?±?0.3 (Pelvetia canaliculata, Phaeophyceae) to 0.8 %?±?0.2 (Porphyra dioica, Rhodophyta). Most common fatty acids were palmitic (16:0), oleic (OLE, 18:1 n-9), α-linolenic (ALA, 18:3 n-3), arachidonic (ARA, 20:4 n-6) and eicosapentaenoic (EPA, 20:5 n-3) acids. Fatty acid profiles were highly variable between and within algal groups; red and brown seaweeds were generally richer in LC-PUFA (e.g. 20:4 n-6 and 20:5 n-3), while high levels of saturated fatty acids such as palmitic acid (16:0) were observed in green species. Most omega-3 PUFA-rich species investigated had a omega-6/omega-3 fatty acid ratio close to 1, which is favourable for human health. The two seasonal sampling times revealed significant differences in total fatty acid and 20:5 n-3 (EPA) contents, with changes depending on species, thus implying varying suitability as potential target species for EPA production. At both times of the year, Palmaria palmata was identified as most promising species as a source of 20:5 n-3 (EPA) amongst all species investigated, with levels ranging from 0.44 to 0.58 % of dry weight in June and November, respectively.     

Characteristics of chimpanzee (Pan troglodytes) ejaculates collected by rectal probe electrostimulation and by artificial vagina

This study compares characteristics of ejaculates collected from 16 adult male chimpanzees using rectal probe electrostimulation (RPE) and from 10 adult male chimpanzees trained to use an artificial vagina (AV). Ejaculate weight, semen volume, and sperm number were significantly lower (P < 0.01) and percentage liquefaction was significantly higher (P < 0.01) in ejaculates collected by RPE. Percentages of motile sperm and of live sperm in semen did not differ significantly between the two collection methods. Total amounts of protein and of α-glucosidase activity were significantly lower (P < 0.01) in seminal fluid from RPE samples. For ejaculates collected by RPE, semen volume correlated positively with protein (r = 0.8640, P < 0.001), fructose (r = 0.6976, P < 0.001), and citrate (r = 0.6976, P < 0.001); sperm number correlated positively with α-glucosidase activity (r = 0.6547, P < 0.001); and protein correlated positively with fructose (r = 0.5906, P < 0.002), citrate (r = 0.5926, P < 0.002) and α-glucosidase activity (r = 0.6006, P < 0.001). For ejaculates collected by AV, semen volume correlated positively with percentage liquefaction (r = 0.6058, P < 0.001), protein (r = 0.8055, P < 0.001), fructose (r = 0.6606, P < 0.001), and citrate (r = 0.8272, P < 0.001); sperm number correlated positively with percentage of motile sperm (r = 0.4196, P 0.004); percentage of motile sperm correlated positively with percentage of live sperm (r = 0.4388, P < 0.002); and, protein correlated positively with fructose (r = 0.6947, P < 0.002) and with citrate (r = 0.5926, P < 0.002). These data show that there is a significant difference in semen parameters and in biochemical parameters of ejaculates obtained by RPE and by AV. © 1995 Wiley-Liss, Inc.     

Genomic and Phenotypic Characterization of Vibrio cholerae Non-O1 Isolates from a US Gulf Coast Cholera Outbreak

Between November 2010, and May 2011, eleven cases of cholera, unrelated to a concurrent outbreak on the island of Hispaniola, were recorded, and the causative agent, Vibrio cholerae serogroup O75, was traced to oysters harvested from Apalachicola Bay, Florida. From the 11 diagnosed cases, eight isolates of V. cholerae were isolated and their genomes were sequenced. Genomic analysis demonstrated the presence of a suite of mobile elements previously shown to be involved in the disease process of cholera (ctxAB, VPI-1 and -2, and a VSP-II like variant) and a phylogenomic analysis showed the isolates to be sister taxa to toxigenic V. cholerae V51 serogroup O141, a clinical strain isolated 23 years earlier. Toxigenic V. cholerae O75 has been repeatedly isolated from clinical cases in the southeastern United States and toxigenic V. cholerae O141 isolates have been isolated globally from clinical cases over several decades. Comparative genomics, phenotypic analyses, and a Caenorhabditis elegans model of infection for the isolates were conducted. This analysis coupled with isolation data of V. cholerae O75 and O141 suggests these strains may represent an underappreciated clade of cholera-causing strains responsible for significant disease burden globally.     

Length-weight relationship of coral reef-associated fishes from Gulf of Mannar and Palk Bay,Southeast Coast of India

Length-weight relationship parameter was estimated for eight coral reef-associated finfish species from the Gulf of Mannar and Palk Bay, Eastern Indian Ocean. Fish samples were collected from the landings of trawlers with a cod-end mesh size of 25–35 mm, operated at a depth ranging from 50–70 m off Tuticorin, Rameswaram and Thondi coasts on monthly basis between June-2018 and July-2019. Fish specimens were sampled by measuring total length (TL) and total weight (TW) with precision to 0.1 cm and 0.1 g respectively. A new maximum total length was recorded for Canthigaster solandri, Centriscus scutatus and Halichoeres stigmaticus from this study.     

Seasonality and Structure of a Macrobenthic Seagrass Community on the Florida Gulf Coast

Macrofauna at two intertidal sites, quantitatively sampled monthly for 25 months, exhibited distinct reoccurring population fluctuations which were poorly correlated with abiotic parameters. Faunal dominants, Laeonereis culveri and Onuphis simoni, showed reciprocal fluctuations. Laeonereis exhibited non-synchronized reproduction and recruitment while Onuphis exhibited synchronized release. Laeonereis peaked before Onuphis while Oniphis fluctuated with the majority of other species. The faunal dominants exhibited one cycle in autumn at both sites but also exhibited a cycle at only one site in spring. Temperature is thus not indicated as causal and detrital input was suggested as the overriding factor. A hypothesis was advanced that Laeonereis is an opportunistic species exploiting fresh detritus, while Onuphis is a detritus feeder utilizing bacterial populations.     

Preservation of necrotizing hepatopancreatitis bacterium (NHPB) by freezing tissue collected from experimentally infected Litopenaeus vannamei

Necrotizing hepatopancreatitis (NHP), a severe disease of penaeid shrimp, is caused by bacteria (NHPB) that have previously been demonstrated to reside in tubular epithelial hepatopancreatic (HP) cells of infected shrimp. There has yet to be a successful in vitro culture method to grow the intracellular organism; therefore, it must be propagated in vivo via transmission from NHPB-infected shrimp to healthy individuals. In our studies, NHPB propagation tanks containing infected shrimp were used to maintain a constant supply of organisms for experiments. In order to develop a method for storing infectious NHPB material for future challenge studies, we collected HP tissue containing NHPB by flash freezing whole, fresh HPs at -80 degrees C for up to 80 d and used it to successfully infect specific pathogen-free Litopenaeus vannamei per os in controlled experiments. HP tissue samples were collected from dead shrimp, and PCR was performed to confirm the presence of NHPB. Our results demonstrate that the infectivity of NHPB in tissue is not altered after being frozen at -80 degrees C when compared to NHPB in fresh tissue. Thus, the continual propagation of NHPB in vivo is not required to assure a source of the infectious agent.     

Saddleback Deformities in Fish Species Collected from the Arabian Gulf Coast of Jubail City,Saudi Arabia

The saddleback syndromes were observed in several commercial fishes obtained from Saudi Arabian coast of the Arabian Gulf. These deformities resulting in missing dorsal spines or rays, sometimes accompanied by a depression in the dorsal profile, were found in ten fish species from seven families. Other morphological abnormalities such as scale disturbances, lateral line deformity, and kyphosis, the presence of a lesion, slight ambicoloration, and supraneural deformities accompanying the saddleback deformity were also observed. The occurrence of similar deformities across such a spectrum of fishes from the same location suggests that the abnormalities were induced by something in the environment common to all these species.