Fish-isolated Naegleria strains and their phylogeny inferred from ITS and SSU rDNA sequences

Effort was made to identify Naegleria strains isolated from organs of fish, using phylogenetic analyses of SSU rDNA and ITS sequences. Eighteen fish-isolated strains studied enlarged substantially the so far available set of Naegleria strains characterized by both molecular markers. The phylogenetic analyses of separate and concatenated SSU rDNA and ITS sequences revealed phylogenetic relationships of strains under study; however, they failed to solve classification of fish-isolated strains into species. The sequence similarity of strain-representatives of Naegleria species as well as data obtained on intragenomic variation of ITS sequences discouraged the authors from the definition of new species. The results of the present study provide evidence of a need to re-evaluate the current practice of setting boundaries between species of the genus Naegleria. Sequences obtained in this study have been deposited in GenBank with accession numbers DQ768714-DQ768743.     

Didymium-like myxogastrids (class Mycetozoa) as endocommensals of sea urchins (Sphaerechinus granularis)

This paper sums up the results of light microscopical, ultrastructural and molecular studies of five strains of amoeboid organisms isolated as endocommensals from coelomic fluid of sea urchins, Sphaerechinus granularis (Lamarck), collected in the Adriatic Sea. The organisms are reported as Didymium-like myxogastrids. Of the life-cycle stages, the attached amoeboids, flagellated trophozoites, cysts and biflagellated swarmers are described. Formation of fruiting bodies was not observed. Although phylogenetic analyses of SSU rDNA sequences indicated a close relationship with Hyperamoeba dachnaya, our sea-urchin strains have not been assigned to the genus Hyperamoeba Alexeieff, 1923. The presence of either one or two flagella reported in phylogenetically closely related organisms and mutually distant phylogenetic positions of strains declared as representatives of the genus Hyperamoeba justify our approach. Data obtained in this study may be useful in future analyses of relationships of the genera Didymium, Hyperamoeba, Physarum and Pseudodidymium as well as in higher-order phylogeny of Myxogastrea.     

Animal models of henipavirus infection: A review

Hendra virus (HeV) and Nipah virus (NiV) form a separate genus Henipavirus within the family Paramyxoviridae, and are classified as biosafety level four pathogens due to their high case fatality rate following human infection and because of the lack of effective vaccines or therapy. Both viruses emerged from their natural reservoir during the last decade of the 20th century, causing severe disease in humans, horses and swine, and infecting a number of other mammalian species. The current review summarises current published data relating to experimental infection of small and large animals, including the natural reservoir species, the Pteropus bat, with HeV or NiV. Susceptibility to infection and virus distribution in the individual species is discussed, along with the pathogenesis, pathological changes, and potential routes of transmission.     

Effect of cutting frequency on species richness and dry matter yield of permanent grassland after grazing cessation

The effect of grazing cessation management has only rarely been investigated. In this study, the influence of cutting regime in the absence of fertilization was examined during 2005–2011 on a semi-natural grassland in Rapotin (Czech Republic) which had been grazed for more than 30 years before the start of the trial. This was conducted as a randomized plot design with five levels of treatment: 4 C – intensive (four cuts per year); 3 C – medium intensive (three cuts per year); 2 C – low intensive (two cuts per year); 1 C – extensive (one cut per year); 0 C – abandoned (control; without any management). Species richness was significantly lower in treatments 1 C and 0 C than in other treatments. Management and year had similar and significant influence on species composition and explained 23% of its variability. Dry matter (DM) yields were found to be lower in three-cut than in two-cut treatments (5.56 t ha^?1 and 6.22 t ha^?1, respectively). In the case of grazing cessation, a suitable compromise of three cuts per year can be recommended to guarantee maintenance of species richness, botanical composition, and appropriate grass forage yield under similar site conditions.     

Nuclearia pattersoni sp. n. (Filosea), a new species of amphizoic amoeba isolated from gills of roach (Rutilus rutilus), and its rickettsial endosymbiont

A new species of amphizoic amoeba, Nuclearia pattersoni sp. n., isolated from gills of Rutilus rutilus L. is described. It is characterised by elongate flattened trophozoites of irregular shape. The longer dimension of their bodies is 13.2 (11.0-15.7) microm. Filopodia radiating mostly from the poles are 2 to 2.5 times longer than the body. The diameter of less frequently observed spherical trophozoites is 8.2-10.8 microm; their filopodia radiate to all directions. Cyst-like stages have shorter pseudopodia that arise from one pole only. The surface of locomotive forms from agar plate cultures has a thin amorphous glycocalyx, while most cells are covered by two layers of extracellular matrix. Mitochondria have flattened cristae, dictyosomes are located in the perinuclear zone. A conspicuous ultrastructural feature of the morphologically similar N. simplex, perinuclear striated band, is not present. Light microscopic and ultrastructural data are completed with the sequence of SSU rRNA gene and phylogenetic analysis including sequences of related taxa. The bacterial endosymbiont found in N. pattersoni type strain RR2G2 is assigned to the genus Rickettsia.     

Experimental inoculation study indicates swine as a potential host for Hendra virus

Hendra virus (HeV) is a zoonotic virus from the family Paramyxoviridae causing fatal disease in humans and horses. Five-week-old Landrace pigs and 5-month-old Gottingen minipigs were inoculated with approximately 10⁷ plaque forming units per animal. In addition to fever and depression exhibited in all infected pigs, one of the two Landrace pigs developed respiratory signs at 5 days post-inoculation (dpi) and one of the Gottingen minipigs developed respiratory signs at 5 dpi and mild neurological signs at 7 dpi. Virus was detected in all infected pigs at 2–5 dpi from oral, nasal, and rectal swabs and at 3–5 dpi from ocular swabs by real-time RT-PCR targeting the HeV M gene. Virus titers in nasal swab samples were as high as 10^4.6 TCID₅₀/mL. The viral RNA was mainly distributed in tissues from respiratory and lymphoid systems at an early stage of infection and the presence of virus was confirmed by virus isolation. Pathological changes and immunohistochemical staining for viral antigen were consistent with the tissue distribution of the virus. This new finding indicates that pigs are susceptible to HeV infections and could potentially play a role as an intermediate host in transmission to humans.     

Human-carnivore coexistence in a traditional rural landscape

Facilitating human-carnivore coexistence is a major conservation concern in human-dominated landscapes worldwide. Useful insights could be gained by studying and understanding the dynamics of human-carnivore coexistence in landscapes in which carnivores and humans have coexisted for a long time. We used a two-pronged approach combining ecological and social data to study coexistence of the brown bear (Ursus arctos) and humans in Transylvania, Romania. First, we surveyed 554 km of walking transects to estimate activity via a bear sign index, namely the proportion of anthills disturbed by bears, and used spatially explicit predictive models to test which biophysical and anthropogenic variables influenced bear activity. Second, we interviewed 86 shepherds and 359 villagers and community representatives to assess conflicts with bears and attitudes of shepherds towards bears. Our interdisciplinary study showed that bears and humans coexisted relatively peacefully despite occasional conflicts. Coexistence appeared to be facilitated by: (1) the availability of large forest blocks that are connected to the source population of bears in the Carpathian Mountains; (2) the use of traditional livestock management to minimize damage from bears; and (3) some tolerance among shepherds to occasional conflict with bears. In contrast, bear activity was unrelated to human settlements, and compensation for livestock losses did not influence people’s attitudes toward bears. Our study shows that coexistence of humans and carnivores is possible, even without direct economic incentives. A key challenge for settings with a discontinuous history of human-carnivore coexistence is to reinstate both practices and attitudes that facilitate coexistence.     

Percutaneous collection of fetal fluids for detection of bovine viral diarrhea virus infection in cattle

OBJECTIVE: To develop a method for percutaneous collection of fetal fluid from cattle in the late stages of gestation and determine whether bovine viral diarrhea virus (BVDV) can be isolated from such fluids. DESIGN: Case series. ANIMALS: 169 pregnant beef cattle. PROCEDURE: Animals were restrained in a squeeze chute, and hair was clipped from a region of the right flank. Pregnancy was confirmed, and fetal fluids were identified by means of abdominal ultrasonography. Fetal fluid was collected with a spinal needle. Virus isolation was performed on fetal fluids, WBC lysates from 160 live calves, and tissues from 12 calves that died or were aborted. Blood samples collected from adult cattle were assayed with an immunoperoxidase monolayer assay. RESULTS: Fourteen animals aborted or delivered premature calves within 3 weeks after fetal fluid collection; however, it could not be determined whether this was a complication of the procedure or attributable to other factors. Results of BVDV isolation from fetal fluid samples were negative for 168 animals. However, a noncytopathic BVDV was isolated from fetal fluid obtained from a 2-year-old heifer; results of the immunoperoxidase assay of serum from this heifer were also positive, and a noncytopathic BVDV was isolated from tissue specimens from a stillborn calf produced by this heifer. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that fetal fluids can be collected percutaneously from cattle in the late stages of gestation and that virus isolation performed on fetal fluids can be used to identify fetuses infected with BVDV in utero. However, safety of the procedure could not be evaluated.     

SPI-1-encoded type III secretion system of Salmonella enterica is required for the suppression of porcine alveolar macrophage cytokine expression

ABSTRACT: Genes localized at Salmonella pathogenicity island-1 (SPI-1) are involved in Salmonella enterica invasion of host non-professional phagocytes. Interestingly, in macrophages, SPI-1-encoded proteins, in addition to invasion, induce cell death via activation of caspase-1 which also cleaves proIL-1β and proIL-18, precursors of 2 proinflammatory cytokines. In this study we were therefore interested in whether SPI-1-encoded type III secretion system (T3SS) may influence proinflammatory response of macrophages. To test this hypothesis, we infected primary porcine alveolar macrophages with wild-type S. Typhimurium and S. Enteritidis and their isogenic SPI-1 deletion mutants. ΔSPI1 mutants of both serovars invaded approx. 5 times less efficiently than the wild-type strains and despite this, macrophages responded to the infection with ΔSPI1 mutants by increased expression of proinflammatory cytokines IL-1β, IL-8, TNFα, IL-23α and GM-CSF. Identical macrophage responses to that induced by the ΔSPI1 mutants were also observed to the infection with sipB but not the sipA mutant. The hilA mutant exhibited an intermediate phenotype between the ΔSPI1 mutant and the wild-type S. Enteritidis. Our results showed that the SPI-1-encoded T3SS is required not only for cell invasion but in macrophages also for the suppression of early proinflammatory cytokine expression.