Rickettsial and mollicute infections in hepatopancreatic cells of cultured Pacific white shrimp (Penaeus vannamei)

Infections by multiple species of bacteria occurred in hepatopancreatic epithelial cells of cultured Pacific white shrimp (Penaeus vannamei). Grossly, hepatopancreases of moribund shrimp were pale white. Light microscopically, hepatopancreatic tubules appeared atrophied and were associated with granulomas. Examination by scanning and transmission electron microscopy revealed heavy cytoplasmic infections by three forms of microorganisms: (1) a rickettsia-like bacterium, (2) a helical form of a mollicute-like bacterium, and (3) a filamentous mollicute-like bacterium. The rod-shaped rickettsia (900 nm long by 300 nm wide) appeared to be free in the cytoplasm and had both a plasma membrane and a cell wall. Neither form of mollicute possessed a cell wall. The helical mollicute was blunt at its wide end (about 260 nm in diameter) where it contained electron-lucent bodies. Helical turns along its tapered axis resembled those of a spiroplasma (the only helical form of mycoplasma in the class Mollicutes) or a spirochete. The helical bacterium did not possess periplasmic flagella characteristic of spirochetes, which lends support to its being a type of spiroplasma. The filamentous mollicute consisted of masses of short, branched filaments 60 nm wide with intermittent spherical dilations and terminal blebs on the branches. The presumed mollicutes have not been reported previously in crustaceans. Each bacterium, or concurrent infections of the bacteria, are pathogenic to cultured shrimp, could impact culture operations and thus deserve more study.     

Assessment of human sperm function after recovery from the female reproductive tract

The physiology and fertile life of human spermatozoa in the female reproductive tract have received little previous attention. A technique was developed for recovering spermatozoa from human cervical mucus at various intervals after artificial insemination. The functions of these cells as measured by penetration of the human zona pellucida and fusion with the zona-free hamster oocytes were examined. Penetration into the zona pellucida was consistently observed when sperm were recovered from 1 to 80 h after insemination. Penetration through the zona into the perivitelline space (PVS) was seen from 1 to 72 h after insemination. Fusion of human sperm with zona-free hamster oocytes was observed from 1 to 48 h after insemination. Motile sperm were recovered 112 and 120 h after insemination with swimming speeds comparable to freshly capacitated spermatozoa. Concentrations of recovered sperm at these longer intervals from insemination were insufficient for sperm-oocyte assays. These studies demonstrate that human spermatozoa aged in vivo may be recovered from cervical mucus for physiologic study, and suggest that the fertile life of human sperm may be 80 h or more.     

Rhesus macaque blastocysts resulting from intracytoplasmic sperm injection of vacuum‐dried spermatozoa*

Background Sperm desiccation is an attractive approach for sperm preservation. In this study, we examined the feasibility and efficiency of intracytoplasmic sperm injection using vacuum‐dried rhesus macaque sperm in CZB medium supplemented with 10% fetal bovine serum. Methods A total of 109 MII oocytes were injected with 69 fresh ejaculated sperm and 40 vacuum‐dried sperm. Results Cleavage occurred in 97% of oocytes injected with fresh, motile sperm and in 88% of oocytes injected with vacuum‐dried sperm. Of the cleaved oocytes, 68% fresh sperm‐injected oocytes and 74% of dried sperm‐injected oocytes developed to the compact morula stage. Blastocyst development was comparable between fresh‐injected (16%) and vacuum‐dried‐injected (17%) oocytes. Differences between treatment groups were not significant. Transmission electron microscopic observation of the blastocysts indicated no detectable differences between fresh sperm and dried sperm‐derived embryos. Conclusions We conclude that vacuum‐dried rhesus macaque sperm are capable of inducing fertilization and development of pre‐implantation embryos when sperm were dried under vacuum and microinjected into normal viable oocytes.     

Parasitic crustaceans as vectors of viruses, with an emphasis on three penaeid viruses

Parasitic crustaceans serve as both hosts and vectors of viruses as well as of parasites and other microbial pathogenic agents. Few of the presumably numerous associations are known, but many can be anticipated. Recently, branchiurans and gnathiid isopods have been documented to host helminths and blood parasites. Because the agents can be observed readily with a microscope, these are better recognized than are the smaller viral, bacterial, and fungal agents. Some agents are harmful to the host of the crustacean parasite and others are not. Viruses probably fit both these categories, since viruses that do not appear pathogenic are often seen in ultrastructural images from a range of invertebrate hosts, including crustaceans. Some viruses have been implicated in causing disease in the host, at least under appropriate conditions. For example, lymphocystis virus may possibly be transmitted to the dermis of its fish hosts by copepods and to the visceral organs by a cymothoid isopod. Similarly, argulid branchiurans seem to transmit the viral agent of spring viremia of carp as well as carp pox, and copepods have been implicated in transmitting infectious hematopoietic necrosis, infectious salmon anemia, and infectious pancreatic necrosis to salmon. Other viruses can be vectored to their hosts through an additional animal. We exposed three viruses, Taura syndrome virus (TSV), white spot syndrome virus (WSSV), and yellowhead virus (YHV), all of which cause mortalities in wild and cultured penaeid shrimps, to crustacean parasites on fish and crabs. Using real-time polymerase chain reaction analysis, we show that TSV in the cyclopoid copepod Ergasilus manicatus on the gill filaments of the Gulf killifish, Fundulus grandis, the acorn barnacle Chelonibia patula on the carapace of the blue crab, Callinectes sapidus, and gooseneck barnacle Octolasmis muelleri on the gills of C. sapidus, can replicate for at least 2 weeks and establish what should be an infective dose. This result was additionally supported by positive in situ hybridization reactions. All three parasites are the first known non-penaeid hosts in which replication occurs. The mean log copy number of WSSV also suggested that replication occurred in E. manicatus. The mean log copy number of YHV gradually decreased in all three parasites and both hosts over the 2-week period. The vector relationships indicate an additional potential means of transmitting and disseminating the disease-causing agents to the highly susceptible and economically valuable penaeid shrimp hosts.     

In vivo uptake of a haem analogue Zn protoporphyrin IX by the human malaria parasite P. falciparum‐infected red blood cells

The cellular traffic of haem during the development of the human malaria parasite Plasmodium falciparum, through the stages R (ring), T (trophozoite) and S (schizonts), was investigated within RBC (red blood cells). When Plasmodium cultures were incubated with a fluorescent haem analogue, ZnPPIX (Zn protoporphyrin IX) the probe was seen at the cytoplasm (R stage), and the vesicle‐like structure distribution pattern was more evident at T and S stages. The temporal sequence of ZnPPIX uptake byP. falciparum‐infected erythrocytes shows that at R and S stages, a time‐increase acquisition of the porphyrin reaches the maximum fluorescence distribution after 60 min; in contrast, at the T stage, the maximum occurs after 120 min of ZnPPIX uptake. The difference in time‐increase acquisition of the porphyrin is in agreement with a maximum activity of haem uptake at the T stage. To gain insights into haem metabolism, recombinant PfHO (P. falciparum haem oxygenase) was expressed, and the conversion of haem into BV (biliverdin) was detected. These findings point out that, in addition to haemozoin formation, the malaria parasite P. falciparum has evolved two distinct mechanisms for dealing with haem toxicity, namely, the uptake of haem into a cellular compartment where haemozoin is formed and HO activity. However, the low Plasmodium HO activity detected reveals that the enzyme appears to be a very inefficient way to scavenge the haem compared with the Plasmodium ability to uptake the haem analogue ZnPPIX and delivering it to the food vacuole.     

Uses,traditional management,perception of variation and preferences in ackee (Blighia sapida K.D. Koenig) fruit traits in Benin: implications for domestication and conservation

Background

Changing lifestyles have recently caused a severe reduction of the gathering of wild food plants. Knowledge about wild food plants and the local environment becomes lost when plants are no longer gathered. In Central Europe popular scientific publications have tried to counter this trend. However, detailed and systematic scientific investigations in distinct regions are needed to understand and preserve wild food uses. This study aims to contribute to these investigations.

Methods

Research was conducted in the hill country east of Graz, Styria, in Austria. Fifteen farmers, most using organic methods, were interviewed in two distinct field research periods between July and November 2008. Data gathering was realized through freelisting and subsequent semi-structured interviews. The culinary use value (CUV) was developed to quantify the culinary importance of plant species. Hierarchical cluster analysis was performed on gathering and use variables to identify culture-specific logical entities of plants. The study presented was conducted within the framework of the master's thesis about wild plant gathering of the first author. Solely data on gathered wild food species is presented here.

Results

Thirty-nine wild food plant and mushroom species were identified as being gathered, whereas 11 species were mentioned by at least 40 percent of the respondents. Fruits and mushrooms are listed frequently, while wild leafy vegetables are gathered rarely. Wild foods are mainly eaten boiled, fried or raw. Three main clusters of wild gathered food species were identified: leaves (used in salads and soups), mushrooms (used in diverse ways) and fruits (eaten raw, with milk (products) or as a jam).

Conclusions

Knowledge about gathering and use of some wild food species is common among farmers in the hill country east of Graz. However, most uses are known by few farmers only. The CUV facilitates the evaluation of the culinary importance of species and makes comparisons between regions and over time possible. The classification following gathering and use variables can be used to better understand how people classify the elements of their environment. The findings of this study add to discussions about food heritage, popularized by organizations like Slow Food, and bear significant potential for organic farmers.     

Margolisianum bulbosum n. gen., n. sp. (Nematoda: Philometridae) from the southern flounder, Paralichthys lethostigma (Pisces: Bothidae), in Mississippi Sound, USA

A new species of a philometrid nematode, Margolisianum bulbosum, is described from the subcutaneous tissue in the mouth (larvigerous females), head (males, ovigerous, and larvigerous females), and eye (preovigerous females) of the southern flounder, Paralichthys lethostigma, from Mississippi Sound. It is placed in a new genus diagnosed by the combination of 8 large, paired but separate cephalic papillae; no inner cephalic papillae; an esophagus with a separate, muscular anterior bulb; a prominent mononuclear esophageal gland; and variable, irregularly distributed cuticular bosses in the females, as well as a vestigial rectum, particularly in larvigerous females. Some female specimens exhibit rows of lateral grooves and longitudinal ridges near the posterior end. Males have 2 small slightly subequal spicules, a barbed gubernaculum, 4 pairs of small cephalic papillae, and a bipartite hypodermal extension within a membranous cuticle on the posterior end. Males, ovigerous females, and larvigerous females appear to be present year round in this sporadic infection in Mississippi.     

First report of a species of Folliculovarium Gu & Shen, 1983 (Bucephalidae: Prosorhynchinae) from European marine waters,with the description of F. mediterraneum n. sp.

Folliculovarium mediterraneum n. sp. is described and figured in detail from the intestine of the Mediterranean moray eel Muraena helena (Muraenidae) in the Scandola Nature Reserve, off Corsica in the Western Mediterranean. This new species closely resembles F. gymnothoracis Gu and Shen, 1983 and F. xishaense Gu and Shen, 1983 from marine fishes off China, but it may differ from both in not having specialised circum-rynchal spines. It differs from the former by having slightly larger eggs and a longer oesophagus, and from F. xishaense by having tegumental spines, a longer oesophagus and a non-related host (an eel rather than a serranid). The status of Folliculovarium, which was recently treated as a genus inquirendum by Overstreet and Curran (2002), is clarified by its multilobed rather than follicular ovary and other features, and remains in the Prosorhynchinae (Bucephalidae).