Identification of optimal regions for phylogenetic studies on VP1 gene of foot-and-mouth disease virus: analysis of types A and O Argentinean viruses

An analysis of the informative content of sequence stretches on the foot-and-mouth disease virus (FMDV) VPI gene was applied to two important viral serotypes: A and O. Several sequence regions were identified to allow the reconstruction of phylogenetic trees equivalent to those derived from the whole VPI gene. The optimal informative regions for sequence windows of 150 to 250 nt were predicted between positions 250 and 550 of the gene. The sequences spanning the 250 nt of the 3' end (positions 400 to 650), extensively used for FMDV phylogenetic analyses, showed a lower informative content. In spite of this, the use of sequences from this region allowed the derivation of phylogenetic trees for type A and type O FMDVs which showed topologies similar to those previously reported for the whole VP1 gene. When the sequences determined for viruses isolated in Argentina, between 1990 and 1993, were included in these analyses, the results obtained revealed features of the circulation of type A and type O viruses in the field, in the months that preceded the eradication of the disease in this country. Type A viruses were closely related to an Argentinean vaccine strain, and defined an independent cluster within this serotype. Among the type O viruses analysed, two groups were distinguished; one was closely related to the South American vaccine strains, while the other was grouped with viruses of the O3 subtype. In addition, a detailed phylogeny for type A FMDV is presented.     

Leishmaniasis in Canis familiaris – a Report About Microvascular and Cellular Architecture of the Infected Spleen

Leishmaniases are a globally widespread group of parasitic diseases from the group of anthropozoonoses. They are caused by an intracellular protozoan parasite that causes a wide spectrum of diseases in humans and dogs worldwide. In fact, the dog is considered one of the most important reservoir. The spleen is one of the several haematopoietic and immunocompetent organs involved. As this organ is a blood filter, the authors looked for the expression of this infection over the microvascular environment and modifications of the spleen cell population related to immunological responses to this parasitic condition.
Tools:  Scanning electronic microscopy over intact tissue and corrosion casts, transmission electronic microscopy, histology and immunohistochemistry (MAC 387 and CD3).
Results:  Our results show three important modifications concerning the microvascular architecture of the spleen when compared to the normal pattern. A striking scarcity of the sinusoidal system sheath that surrounds the central arteries of the white pulp, a huge development of pulp venules and veins, differentiation of typical features of high endothelial venular cells.
Remarks:  According to our results it seems that independent of the virulence of the parasite involved and the kind of immunity prevalent in a particular host, the spleen develops blood dynamic conditions that allows a sluggish blood flow so that cells involved in immunological processes can proliferate and differentiate and it also contributes to the trapping of lymphocytes to the area through the differentiation of typical characteristics of HEV endothelial cells.     

Use of touch-down polymerase chain reaction to enhance the sensitivity of Mycobacterium bovis detection.

The confirmatory diagnosis of Mycobacterium bovis (M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This study describes the development of a single-tube touch-down polymerase chain reaction (PCR) protocol for the detection of M. bovis using primers that target the IS6110 element. Spiked water and milk as well as routine diagnostic samples (milk and nasal swabs) from M. bovis-positive cattle were tested. This protocol allows the rapid and sensitive detection of M. bovis in bovine samples by enhancing the sensitivity of standard PCR amplification.     

Pneumatosis cystoides intestinalis in a rabbit doe (Oryctolagus cuniculus).

Pneumatosis cystoides intestinalis (PCI) is an infrequent condition of animals characterized by the existence of numerous thin-walled, gas-filled cystic structures within the intestinal wall and adjacent lymph nodes. Microscopically, the cystic structures appear to be dilated lymphatics located in the lamina propria, submucosa, muscularis, subserosa, mesentery, and mesenteric lymph nodes. This report describes a case of pneumatosis cystoides intestinalis in a rabbit doe from an organic farm where 20 rabbit does were fed ad libitum with a natural diet consisting of whole barley, pea beans, alfalfa hay, and a pelleted vitamin-mineral blend. A combination of nutritional, bacterial, and other factors are hypothesized as possible predisposing factors in the development of PCI.     

Use of avidity enzyme-linked immunosorbent assay and avidity Western blot to discriminate between acute and chronic Neospora caninum infection in cattle.

Avidity serological tests (avidity enzyme-linked immunosorbent assay [ELISA] and avidity Western blot) were developed and used to differentiate between acute (primary infection, reinfection, and recrudescence) and chronic Neospora caninum infection in cattle. In addition, the pattern of immunoglobulin G (IgG) avidity maturation against different specific antigens of N. caninum tachyzoites was studied. Sequential serum samples were collected from cattle naturally and experimentally infected with N. caninum. Four groups of experimentally infected cattle were included in the study and were representative of primary infection, reinfection, chronic infection, and noninfection. Serum samples were also collected from naturally infected cattle classified into nonaborting and aborting cows on the basis of clinical findings and serological profiles, and a third group composed of seronegative cows that seroconverted during the course of the experiment. All samples were tested by avidity ELISA and avidity Western blot. The IgG avidity ELISA allowed the discrimination between primary and chronic infection because all experimentally primary-infection cows showed low avidity indexes at week 4 postinfection (p.i.) compared with the high avidity values found at week 20 postinfection. However, this test did not allow the discrimination of reinfection or recrudescence from chronic infection. Regarding IgG avidity Western blot results, no antigenic markers correlating with acute (primary infection, recrudescence, and reinfection) or chronic infection were recognized. However, the 17-kD immunodominant antigen was mostly responsible for high avidity values obtained by avidity ELISA because it was intensively recognized by high-avidity antibodies in all chronically infected animals after urea treatment.     

Isolation of equine herpesvirus-2 from the lung of an aborted fetus.

This study describes the isolation of equine herpesvirus-2 (EHV-2) from the lung of an aborted equine fetus in Argentina. The isolated virus was confirmed as EHV-2 by indirect immunofluorescence using a rabbit anti-EHV-2 polyclonal antiserum and by virus-neutralization test using an equine polyclonal antibody against EHV-2. Restriction endonuclease DNA fingerprinting with BamHI also confirmed the identity of the virus as EHV-2. Furthermore, viral nucleic acid was detected by polymerase chain reaction from the original lung sample and from the DNA obtained from cells infected with the virus isolate. This work constitutes the first reported isolation of EHV-2 from an aborted equine fetus. The presence of EHV-2 in the lung of the aborted fetus would indicate that this virus is capable of crossing the placental barrier. However, no cause-effect relationship was established between the EHV-2 isolate and the abortion.     

An in vitro study of hematopoietic progenitor cells from peripheral blood of rabbits

The purpose of this study was to isolate and cultivate a subpopulation of pluripotent stem cells (PSCs) from the peripheral blood of rabbits, which are frequently used in veterinary research as an animal model. Pluripotent stem cells, as described in human beings, are fibroblast-like cells that exhibit a CD34 marker, specific from other hematopoietic stem cells. Commonly used human commercial media has been researched for culturing rabbit PSCs. These findings allow us to contemplate the direct application of this simple and standardized methodology in several areas of study, such as of the pharmacological effect of many drugs on hematopoietic cells, veterinary practice, and even the study of new strategies in cellular therapy for some human diseases.     

Nutritional Enhancement of Biocontrol Activity of Candida sake (CPA-1) against Penicillium expansum on Apples and Pears

Pome fruits are poor in nitrogenous compounds and the addition of nitrogen can improve colonisation of the fruits by antagonists. Twenty-two nitrogenous compounds were evaluated for their effect on Candida sake (CPA-1) growth in vitro. Ten compounds that induced greater growth were applied with the antagonist to wounded fruits to evaluate their effect on enhancing control of Penicillium expansum. Calcium chloride and 2-deoxy-D-glucose were also tested. L-serine and L-aspartic acid enhanced biocontrol by C. sake against P. expansum on apples. On apples and pears, ammonium molybdate, calcium chloride and 2-deoxy-D-glucose improved the capacity of the antagonist to control P. expansum. The addition of ammonium molybdate at 1 mM allowed C. sake to be used on apples and pears at a lower concentration without diminishing control. Similar results were observed with the addition of calcium chloride to the antagonist. 2-deoxy-D-glucose at 6 and 18 mM enhanced biocontrol on pears by over 81%, but on apples the improvement of biocontrol was observed only at 6 mM. In cold storage, the combination of ammonium molybdate and C. sake completely eliminated the incidence of blue mould on pears, and reduced its severity and incidence by more than 80% on apples.     

Using chlorophyll fluorescence induction for a quantitative detoxification assay with metribuzin and chlorotoluron in excised wheat (Triticum aestivum and Triticum durum) leaves

Chlorophyll fluorescence induction was used as a probe to detect herbicide detoxification in tolerant or susceptible wheat cultivars. Experimental conditions have been carefully examined for establishing detoxification kinetics of chlorotoluron and metribuzin, two photosystem-II-inhibiting herbicides. After a root treatment, leaves were cut, placed in glass tubes and maintained in the dark. The fluorescence induction rise was examined repeatedly and detoxification kinetics were established from these data for the same position on the individual leaves. The herbicide-dependent fluorescence rise decreased within hours in chlorotoluron-tolerant but not in susceptible Triticum aestivum cultivars. In contrast, no significant reversion could be detected after metribuzin application in both tolerant and susceptible cultivars of Triticum durum. Near the fluorescence-determined half-inhibition of photosystem II, linear detoxification kinetics were obtained in individual leaves, thus providing an accurate measurement of relative detoxification rates.