Common variable immunodeficiency in three horses with presumptive bacterial meningitis

Three adult horses were evaluated for signs of musculoskeletal pain, dullness, ataxia, and seizures. A diagnosis of bacterial meningitis was made on the basis of results of CSF analysis. Because primary bacterial meningitis is so rare in adult horses without any history of generalized sepsis or trauma, immune function testing was pursued. Flow cytometric phenotyping of peripheral blood lymphocytes was performed, and proliferation of peripheral blood lymphocytes in response to concanavalin A, phytohemagglutinin, pokeweed mitogen, and lipopolysaccharide was determined. Serum IgA, IgM, and IgG concentrations were measured by means of radial immunodiffusion, and serum concentrations of IgG isotypes were assessed with a capture antibody ELISA. Serum tetanus antibody concentrations were measured before and 1 month after tetanus toxoid administration. Phagocytosis and oxidative burst activity of isolated peripheral blood phagocytes were evaluated by means of simultaneous flow cytometric analysis. Persistent B-cell lymphopenia, hypogammaglobulinemia, and abnormal in vitro responses to mitogens were detected in all 3 horses, and a diagnosis of common variable immunodeficiency was made.     

Serologic and molecular characterization of Anaplasma species infection in farm animals and ticks from Sicily

Although Anaplasma marginale was known to be endemic in Italy, the diversity of Anaplasma spp. from this area have not been characterized. In this study, the prevalence of Anaplasma spp. antibodies in randomly selected farm animals collected on the island of Sicily was determined by use of a MSP5 cELISA for Anaplasma spp. and an immunofluorescence test specific for Anaplasma phagocytophilum. Genetic variation among strains of Anaplasma spp. from animals and ticks was characterized using the A. marginale msp1alpha and the Anaplasma spp. msp4 genes. Eight species of ticks were collected and tested by PCR. Seropositivity for Anaplasma spp. and A. phagocytophilum was detected in bovine and ovine samples. All the donkeys were seropositive for A. phagocytophilum but not for Anaplasma spp. Four A. marginale genotypes were identified by msp4 sequences from bovine and tick samples. Two new genotypes of Anaplasma ovis were characterized in sheep. The sequences of A. phagocytophilum from three donkeys proved to be identical to the sequence of the MRK equine isolate from California. Six A. marginale genotypes were found in cattle and one tick using the A. marginale msp1alpha sequences. All genotypes had four repeated sequences in the N-terminal portion of the MSP1a, except for one that had five repeats. The Italian strains of A. marginale contained three repeat sequences that were not reported previously. Definition of the diversity of Anaplasma spp. in Sicily reported, herein is fundamental to development of control strategies for A. marginale, A. ovis and A. phagocytophilum in Sicily.     

Papular dermatitis due to Leishmania spp. infection in dogs with parasite-specific cellular immune responses

Papular dermatitis due to Leishmania spp. infection was diagnosed in three boxers and two Rottweilers with Leishmania-specific cellular immunity. Diagnosis was based on histological and immunohistochemical examination of papules in four dogs and on cytological examination in one dog. Serum protein electrophoresis was within reference ranges and low antibody levels to Leishmania infantum were detected. Delayed-type hypersensitivity (DTH) reaction to leishmanin was evaluated before treatment in three dogs with positive results. After meglumine antimoniate therapy for 3 to 4 weeks and allopurinol treatment for 6 to 10 months, all dogs were clinically normal, had positive DTH reactions to leishmanin and reduced antibody titres. In conclusion, we suggest that this previously unreported cutaneous presentation of canine leishmaniosis appears to be associated with specific immunocompetence and, consequently, with a favourable prognosis.     

Identification of gender-regulated genes in Ancylostoma braziliense by real-time RT-PCR

Ancylostoma braziliense belongs to the family Ancylostomatidae and infects cats and dogs in various parts of the tropical world. It is also a zoonotic parasite causing cutaneous larva migrants in humans. There are very few, either biological or molecular, studies of this species. In this study, differential display was used to identify differentially expressed genes in male and female A. braziliense. Nineteen new sequences were identified and examined by real-time RT-PCR to confirm male-female specificity. Ten were more expressed in males, while two were more expressed in females. Molecules shown to be important in other host-parasite relationships were also found in this study.     

Molecular diversity of Brazilian strains of porcine circovirus type 2 (PCV-2)

Porcine circovirus 2 (PCV-2) is associated with a broad range of syndromes. In this study, eight pig tissue samples from two Brazilian states were analyzed using six PCR primer pairs amplifying a 1705-bp fragment of the PCV-2 genome. The NJ distance-based method was used for the phylogenetic analysis with the eight field strains herein, 15 GenBank sequences and using PCV-1 as an out-group. This yielded two major clusters (A and B) for this viral species, with the Brazilian strains segregating with European and Asian sequences. Nucleotide identity was 99.7 to 100% among the sequences. This information can be used in further studies of pathogenesis related to PCV-2 in Brazil.     

Disentangling the trophic interactions between American bullfrogs and native anurans: Complications resulting from post‐metamorphic ontogenetic niche shifts

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Endogenous and exogenous polyamines in the organogenesis in Curcuma longa L.

The present work evaluated the development of different Curcuma longa L. explants (leaves basis, root tips and ancillary buds from rhizome) stimulated by exogenous polyamines, combined with naphtalen-acetic acid (NAA) or with 6-benzyl-aminopurine (BAP), to produce callus and its subsequent differentiation. The explants, isolated from field plants, were previously subjected to a basic cleaning method and were inoculated onto Murashige and Skoog culture medium (MS) [Murashige, T.S., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum 15, 473–497] supplemented with NAA (2.0 mg L⁻¹). Buds were subjected to different treatments, with or without 5.0 and 10.0 mmol L⁻¹ exogenous polyamines (mixture of putrescine:spermine:spermidine, 1:1:1) combined with NAA. The calluses obtained were transferred into the same medium, supplemented with the mixture of polyamines combined with BAP, in order to induce plant differentiation. For C. longa, buds were the most efficient explants for callus induction (p < 0.05). The application of exogenous polyamines (5.0 and 10.0 mmol L⁻¹) produced the most developed callus, with numerous roots. The medium supplemented with 10 mmol L⁻¹ polyamine mixture, combined with BAP, induced good regeneration, producing vigorous plants and excellent shoot formation.Polyamines addition promoted the formation of callus, roots and leaves, representing an important factor in the determination of indirect organogenesis in C. longa L., and putrescine content may be considered a valuable marker of the differentiation process in this specie, as well as the enzyme peroxidase.     

Increased nitrogen-use efficiency of a short-rotation poplar plantation in elevated CO(2) concentration

We estimated nitrogen (N) use by trees of three poplar species exposed for 3 years to free air CO(2) enrichment (FACE) and determined whether the CO(2) treatment affected the future N availability of the plantation. Trees were harvested at the end of the first 3-year rotation and N concentration and content of woody tissues determined. Nitrogen uptake of fine roots and litter was measured throughout the first crop rotation. The results were related to previously published variations in soil N content during the same period. We estimated retranslocation from green leaves and processes determining N mobilization and immobilization, such as mineralization and nitrification, and N immobilization in litter and microbial biomass. In all species, elevated CO(2) concentration ([CO(2)]) significantly increased nitrogen-use efficiency (NUE; net primary productivity per unit of annual N uptake), decreased N concentration in most plant tissues, but did not significantly change cumulative N uptake by trees over the rotation. Total soil N was depleted more in elevated [CO(2)] than in ambient [CO(2)], although not significantly for all soil layers. The effect of elevated [CO(2)] was usually similar for all species, although differences among species were sometimes significant. During the first 3-year rotation, productivity of the plantation remained high in the elevated [CO(2)] treatment. However, we observed a potential reduction in N availability in response to elevated [CO(2)].