Molecular and morphological delineation of <Emphasis Type="Italic">Longidorus poessneckensis</Emphasis> Altherr, 1974 (Nematoda: Dorylaimida)

The plant–parasitic nematode Longidorus poessneckensis from the Czech Republic was morphologically and molecularly characterised. Molecular analyses were carried out using mitochondrial DNA (cytochrome c oxidase subunit 1—cox1) and ribosomal DNA (ITS2—second internal transcribed spacer, 18S gene and D2/D3 expansion segments of the 28S gene), which were amplified and sequenced. Phylogenetic relationship of L. poessneckensis with three morphologically closely related species, i.e. L. macrosoma, L. helveticus and L. uroshis, was inferred by using maximum likelihood and maximum parsimony methods, with a female of Xiphinema diversicaudatum and a bivulval female of X. vuittenezi as outgroups. All multiple alignments yielded similar basic trees supporting the uniqueness of L. poessneckensis and the validity of the four Longidorus species identified using morphological characters. Phylogenetic analyses revealed that L. poessneckensis is more closely related to L. macrosoma and L. helveticus than to L. uroshis. High inter-population diversity (19%) was observed across the cox1 gene between two populations of L. poessneckensis.     

Cryptosporidiumparvum oocysts in seawater clams (Chameleagallina) in Italy

Bivalves filter large volumes of water and can concentrate organisms which are pathogenic for humans and animals. Our aim was to evaluate the presence of Cryptosporidium spp. in clams from the Adriatic coast (Abruzzo region) and genetically characterize the oocysts isolated from the clams. From March to July 2003, 960 specimens of clams (Chamelea gallina) present in nature were collected at 500 m from the Tordino, Tronto, Vibrata and Vomano river mouths on the Adriatic sea. The haemolymph and tissues were extracted from the specimens (240 per river mouth) after the specimens had been identified, measured and weighed (live weight). Immunofluorescence tests (IFA) were performed on pools (n = 32) of samples and oocysts of Cryptosporidium spp. were detected in 23 pools of C. gallina. To identify the Cryptosporidium species, all the pools IFA-positive were tested by a PCR assay specific for the Cryptosporidium outer wall protein (COWP) gene. Positive amplicons then were sequenced and analysed. Two pools of clams were positive for Cryptosporidium parvum Genotype 2 (the “bovine” i.e. zoonotic genotype). This is the first time that C. parvum was found in clams from the Adriatic sea in Italy and the case might be of public health importance.     

Efficacy of moxidectin injectable and pour-on formulations in a pilot control program against bovine hypodermosis in Southern Italy

Bovine hypodermosis is a myiasis caused by Hypoderma bovis and Hypoderma lineatum (Diptera, Oestridae) larvae, which has a severe economic impact on the livestock industry. Though myiasis is widespread throughout Italy, no nationwide eradication program has ever been planned, unlike in other European Countries. With a view to setting up a national control program, a pilot study was carried out in Southern Italy on 9939 cattle bred in an area with a high prevalence of cattle hypodermosis, using moxidectin 0.5% pour-on (Cydectin, Fort Dodge) and 1% injectable (Cydectin, Fort Dodge) formulations. At the recommended dosage, moxidectin displayed efficacy levels of 99.9% in the pour-on and 100% in the injectable formulation, whereas the microdose (1 mg per head regardless of body weight) was less effective (65.7%). This trial contributed to a significant reduction in infestation rates in the study area and represented the first step through which a national program for eradicating warble fly infestation in Italy.     

Typing of Haemophilus parasuis strains by PCR-RFLP analysis of the tbpA gene

On the basis of a species-specific PCR assay, a RFLP analysis for typing of Haemophilus parasuis strains was developed and evaluated. Amplification was based on the gene tbpA, encoding a transferrin-binding protein. RFLP analysis of the 1.9-kb tbpA-amplicon using TaqI, AvaI and RsaI endonucleases produced 12 different patterns for the reference strains of the 15 known H. parasuis serovars, and showed a high heterogeneity (33 RFLP groups) for 101 H. parasuis clinical isolates tested. The sensitivity, typeability (100% versus 65% for immunodiffusion), high degree of discrimination (0.93 versus 0.84 for immunodiffusion), simplicity and low cost per test make this PCR-RFLP assay a useful method for typing H. parasuis and, therefore, for studying the epidemiology of outbreaks of Gl?sser's disease.     

Simultaneous serological evidence of Actinobacillus pleuropneumoniae, PRRS, Aujeszky's disease and influenza viruses in Spanish finishing pigs

A total of 198 pigs with tachypnoea and temperature >/= 40 degrees C were selected on a Spanish finishing unit, and their sera were examined for antibodies to Actinobacillus pleuropneumoniae (App), porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky' disease virus (ADV), and swine influenza virus (SIV). Eighty-nine point nine per cent of the pigs were seropositive to App, 88.6 per cent to PRRS, 73.0 per cent to ADV, and 30.6 per cent to SIV. Thirty-one pigs (15.6 per cent) were seropositive for App, PRRSV, ADV and SIV, and only one (0.5 per cent) was seronegative for all. Statistical association was assessed for dual infections but it was not found in any case (P > 0.05). Other parameters (dyspnoea, nasal discharge and coughing) were also recorded, and no significant associations between them and the presence of antibodies against any of the four infections was found.     

Quantifying by monoclonal antibodies of specific IgG, IgM and IgA in the serum of minipigs experimentally infected with Actinobacillus pleuropneumoniae.

Fifteen minipigs were infected intratracheally with three different doses (10(8), 10(5) or 5 x 10(3) colony forming units) of the reference strains of serotypes 2 or 4 of Actinobacillus pleuropneumoniae, and three remained as controls. The titre of specific IgG, IgM and IgA in the serum was measured weekly for 15 weeks with an indirect ELISA using monoclonal antibodies specific for each isotype. IgG attained the highest titres, IgM lower and IgA the lowest, being only detected in four animals. Serotype 4 evoked significantly higher titres than serotype 2 (P < or = 0.01). In general the highest IgG titres were attained at four to six weeks after infection. Some of the minipigs were reinfected after seven weeks but this evoked an increased titre in only two instances.     

Electron Microscopic Observation on the Structure of a Freshwater Teleost Dermis with Special Reference to the Dermo-epidermal Junction

An electronmicroscopic study of the dermis of the ventral skin of a fresh water teleost ( Pimelodus maculatus ) was examined with standard techniques. It was demonstrated that the dermal organization consists of a well developed basal membrane underlying the epidermis, and two collagenous layers (stratum compactum most superficially and stratum spongiosum, deep to it) which are separated by a layer of elastic connective tissue. Within the superficial layer melanocytes and iridophores were demonstrated.     

Recombinant K39 dipstick immunochromatographic test: a new tool for the serodiagnosis of canine leishmaniasis.

The spread of human leishmaniasis has prompted the scientific community to study dogs as reservoirs for Leishmania infantum. Canine leishmaniasis (CanL) is widespread in the Mediterranean area with a prevalence of up to 50%. The first step toward controlling the disease is to monitor its distribution, mainly in stray dogs. The validity of a recombinant K39 (rK39) dipstick test, commercially available for the serodiagnosis of human leishmaniasis, was evaluated using sera from 165 dogs selected on the basis of positive or negative lymph node smears at parasitological examination. The results were compared with the indirect fluorescent antibody test (IFAT) (cutoff 1:80). Sera from a group of dogs with other diagnosed diseases but negative for leishmaniasis were also tested to evaluate any cross-reactivity. Various procedures were used for testing whole blood samples. The relative specificity of the rK39 dipstick and IFAT was 100% (97 of 97) and 98.97% (96 of 97), whereas the relative sensitivity was 97.06% (66 of 68) and 98.53% (67 of 68), respectively. The results of the dipstick and IFAT corresponded except for 2 sera (k = 0.987). This data confirm the usefulness of rK39 antigen for diagnosing CanL both in symptomatic and asymptomatic dogs. The rK39 dipstick proved to be a rapid, sensitive, and specific test that may be very useful in the field for large-scale screening and also in veterinary practice, requiring minimal equipment and operator expertise.