New fossil fish <Emphasis Type="Italic">Musculopedunculus micklichi</Emphasis> (Trichiuroidei) from the lower Oligocene of Germany

The new fossil fish Musculopedunculus micklichi (type locality: the lower Oligocene slate sediments to the south of Heidelberg, FRG) is described from an incomplete skeleton impression. The diagnostic characters are the powerful and relatively deep caudal peduncle with the hemal and, probably, neural processes of anterior preural centra strongly enlarged, located at an angle of about 60°, as well as a compressed body, high number of vertebrae (probably, about 80), the dorsal fin probably slightly differentiated into the spiny and soft parts, and a well-developed fronto-occipital sagittal crest. On the basis of these and other features, the fossil taxon does not conform to the known Euzaphlegidae, Gempylidae, and Trichiuridae and is recognized as the new family Musculopedunculidae probably belonging to the Trichiuroidae.     

Materials for the revision of the family Caristiidae (Perciformes): 1. Description of Paracaristius heemstrai gen. et sp. nov.

All representatives of the oceanic mesobenthopelagic family Caristiidae are rare. This family comprises of two genera and six species, but the status of these taxa remains indistinct. On the basis of examinations of collections from both the Atlantic and Pacific Ocean, and from published data a new genus Paracaristius is established. It differs by a wide suborbital space, the upper jaw is fully covered with suborbitalia, and there is a weak dentition of the roof of the mouth cavity. Within this genus two species are attributed, P. maderensis (Maul, 1949), initially attributed to the genus Caristius (Gill et Smith, 1905), and P. heemstrai sp. nova.     

Four novel PYFs: members of NPY/PP peptide superfamily from the eyestalk of the giant tiger prawn Penaeus monodon

An immunocytochemical method was used for localization of pancreatic polypeptide (PP) immunoreactive substances in the eyestalk of Penaeus monodon using anti-C-terminal hexapeptide of PP (anti-PP6) antiserum. Approximately 200 neuronal cell bodies were recognized in the ganglia between the medulla interna (MI) and medulla terminalis (MT) and surrounding MT in conjunction with the neuronal processes in medulla externa (ME), MI, MT and sinus gland. About half of the PP immunoreactive neurons were also recognized by a combination of three monoclonal antibodies raised against FMRFamide-like peptides. Isolation of the PP immunoreactive substances from the eyestalk was performed using 7500 eyestalks extracted in methanol/acetic acid/water (90/1/9) followed by five to six steps of RP-HPLC separation. Dot-ELISA with anti-PP6 antiserum was used to monitor PP-like substances in various fractions during the purification processes. Four new sequences of one hexapeptide; RARPRFamide, and three nonapeptides; YSQVSRPRFamide, YAIAGRPRFamide and YSLRARPRFamide were identified, and named as Pem-PYF1-4 due to their structural similarity to the PYF found in squid Loligo vulgaris. Each of the new peptides shares four to seven common residues with the C-terminus of the squid PYF and with the NPFs found in other invertebrates. The NPY/PP superfamily as well as the FMRFamide peptide family may be present throughout vertebrates and invertebrates.     

Specific monoclonal antibodies raised against Taura syndrome virus (TSV) capsid protein VP3 detect TSV in single and dual infections with white spot syndrome virus (WSSV)

The gene sequence encoding VP3 capsid protein of Taura syndrome virus (TSV) was cloned into pGEX-6P-1 expression vector and transformed into Escherichia coli BL21. After induction, recombinant GST-VP3 (rVP3) fusion protein was obtained and further purified by electro-elution before use in immunizing Swiss mice for production of monoclonal antibodies (MAb). One MAb specific to glutathione-S-transferase (GST) and 6 MAb specific to VP3 were selected using dot blotting and Western blotting. MAb specific to VP3 could be used to detect natural TSV infections in farmed whiteleg shrimp Penaeus vannamei by dot blotting and Western blotting, without cross reaction to shrimp tissues or other shrimp viruses, such as white spot syndrome virus (WSSV), yellow head virus (YHV), monodon baculovirus (MBV) and hepatopancreatic parvovirus (HPV). These MAb were also used together with those specific for WSSV to successfully detect TSV and WSSV in dual infections in farmed P. vannamei.     

Differentiation among the Vibrio cholerae serotypes O1, O139, O141 and non-O1, non-O139, non-O141 using specific monoclonal antibodies with dot blotting

Seven different monoclonal antibodies (MAbs) specific to only Vibrio cholerae were produced using a combination of five representative serotypes of V. cholerae for immunization. The first three MAbs (VC-93, VC-82 and VC-223) were specific to the V. cholerae serogroup O1 with different avidity for the serotypes O1 Inaba and O1 Ogawa. The fourth and the fifth MAbs were specific to V. cholerae O139 (VC-812) or O141 (VC-191) serogroups, respectively. The sixth MAb (VC-26) bound to all three serogroups of V. cholerae. The seventh MAb (VC-63) bound to all twenty five isolates of V. cholerae used in this study. None of the seven MAbs showed cross-reactivity with other Vibrio spp. or closely-related V. cholerae species, V. mimicus or other gram-negative bacteria. The eighth MAbs (VC-201) specific to almost all Vibrio spp. was also obtained. In dot blotting, these MAbs can be used to detect a diluted pure culture of V. cholerae in solution with a sensitivity range of from 10⁵ to 10⁷ CFU ml^− 1. However, the detection capability could be improved equivalent to that of PCR technique after preincubation of samples in alkaline peptone water (APW). Thus, these MAbs constitute convenient immunological tools that can be used for simple, rapid and simultaneous direct detection and differentiation of the individual serotypes of V. cholerae in complex samples, such as food and infected animals, without the requirement for bacterial isolation or biochemical characterization.     

A comparative description and distribution of the flying fishes—Cypselurus poecilopterus, C. simus, and C. callopterus, sorted out into the species group of spotwing species of the subgenus Poecilocypselurus

The three species of flying fishes—Indo-Pacific Cypselurus poecilopterus, Central Pacific C. simus, and Eastern Pacific C. callopterus form a particular species group within the subgenus Poecilocypselurus. These species rather weakly differ in the body height, number of predorsal scales and number of gill rakers. The sculls of these three species are very much alike; there are just very small differences in the proportions of some bones and the number of skull lateral line canals and pores. It could be imagined that C. poecilopterus has appeared in the Indo-Malayan Archipelago area and thence extended in all directions (up to the coasts of the Eastern Africa, Northern and Eastern Australia, New Guinea and adjacent islands, the western islands of Polynesia and the waters of Japan). C. callopterus inhabiting warm waters of the Eastern Pacific and C. simus whose area is situated in the central part of the Pacific Ocean are seemingly derived from this species.     

Generation of monoclonal antibodies specific to Hepatopancreatic parvovirus (HPV) from Penaeus monodon

Hepatopancreatic parvovirus (HPV) was isolated from the hepatopancreas (HP) of slow growth Penaeus monodon by urografin gradient centrifugation. The presence of HPV in the fraction was monitored by PCR and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Only 1 major 54 kDa protein band was observed in the strong PCR-positive fractions used to immunize mice for monoclonal antibody production. After cell fusion, the first step in selecting specific antibodies was performed by dot-blot assay with purified HPV viral particles. The second screening step was carried out using Western blots of purified HPV proteins and immunohistochemistry of HPV-infected HP tissue. Four monoclonal antibodies were isolated; these bound to the 54 kDa protein in Western blots and to intranuclear inclusion bodies in tubule epithelial cells of HPV-infected prawn tissue by immunohistochemistry. None of the antibodies showed cross-reactivity either to uninfected shrimp tissue or to other shrimp viruses tested. These reagents have potential for use in developing a highly sensitive immunoassay such as sandwich ELISA or a convenient kit for detection of HPV infection.