Grazia Carelli

GENETIC DIVERSITY OF ANAPLASMA SPECIES MAJOR SURFACE PROTEINS AND IMPLICATIONS FOR ANAPLASMOSIS SERODIAGNOSIS AND VACCINE DEVELOPMENT

Abstract The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes several pathogens of veterinary and human medical importance. An understanding of the diversity of Anaplasma major surface proteins (MSPs), including those MSPs that modulate infection, development of persistent infections, and transmission of pathogens by ticks, is derived in part, by characterization and phylogenetic analyses of geographic strains. Information concerning the genetic diversity of Anaplasma spp. MSPs will likely influence the development of serodiagnostic assays and vaccine strategies for the control of anaplasmosis.

Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Babesia bovis

ABSTRACT A previously developed competitive enzyme-linked immunosorbent assay (cELISA) based on a species-specific, broadly conserved, and tandemly repeated B-cell epitope within the C terminus of rhoptry-associated protein 1 of Babesia bovis was refined and validated for use internationally. Receiver operating characteristic analysis revealed an assay with a specificity and positive predictive value of 100% and a sensitivity of 91.1%, with various negative predictive values depending on the level of disease prevalence. The cELISA was distributed to four different laboratories, along with a reference set of 100 defined bovine sera, including known-positive, known-negative, and field samples. Pairwise concordances among the four laboratories ranged from 94% to 88%. Analysis of variance of the resulting optical densities and a test of homogeneity indicated no significant difference among the laboratories. Overall, the cELISA appears to have the attributes necessary for international application.

Duplex real-time polymerase chain reaction for simultaneous detection and quantification of Anaplasma marginale and Anaplasma centrale.

Anaplasma marginale and Anaplasma centrale are rickettsial pathogens responsible for acute disease and mild infections, respectively, in cattle herds. A duplex real-time polymerase chain reaction (PCR) assay with probes labeled with different fluorophores was developed for simultaneous detection and quantification of A. marginale and A. centrale DNA in bovine blood samples. The assay was able to detect as few as 101 and 102 DNA copies for A. marginale and A. centrale, respectively, with optimal specificity and reproducibility. Analysis by real-time and nested PCR carried out on 54 samples previously tested by reverse line blot hybridization showed that the established duplex real-time PCR assay can detect and quantify the 2 Anaplasma spp., even if present simultaneously in the same blood samples. Such an assay could be used in pathogenesis studies on bovine acute anaplasmosis.

Acute-Phase Proteins in Dogs Naturally Infected with Leishmania infantum During and After Long-term Therapy with Allopurinol

Sasanelli, M., Paradies, P., de Caprariis, D., Greco, B., De Palo, P., Palmisano, D. and Carelli, G., 2007. Acute-phase proteins in dogs naturally infected with Leishmaniainfantum during and after long-term therapy with allopurinol. Veterinary Research Communications, 31(Suppl. 1), 335–338

Retrospective study of hemoparasites in cattle in southern Italy by reverse line blot hybridization.

ABSTRACT Tick-borne diseases are widespread in tropical and temperate regions and are responsible for important economic losses in those areas. In order to assess the presence and prevalence of various pathogens in southern Italy, we retrospectively analyzed cattle blood samples collected for a previous study in 2000 using reverse line blot (RLB) hybridization. The study had been carried out in three regions of southern Italy on 1,500 randomly selected and apparently healthy adult cattle. RLB showed that 43.7% of the cattle were positive for nine different species of hemoparasites with either a single infection or a mixed infection. Theileria buffeli was the most common species found, being present in 27.3% of the animals, followed by Anaplasma marginale in 18.1%, Anaplasma centrale in 13.8%, Babesia bigemina and Anaplasma bovis in 4.2%, Anaplasma phagocytophilum in 1.7%, Babesia bovis in 1.6%, Babesia major in 0.2% and Babesia divergens in 0.1%. Complete blood counts showed different degrees of anemia in 363 animals (24.2%) and of these, 169 were RLB-positive for at least one pathogen. Among the ticks that were collected from the cattle, the following species were identified: Rhipicephalus bursa, Ixodes ricinus, Hyalomma marginatum, Boophilus annulatus, Dermacentor marginatus and Haemaphysalis (sulcata, parva, inermis and punctata). The results obtained confirmed the spread of endemic tick-borne pathogens in the regions studied.

First Report of Bovine Anaplasmosis Caused by Anaplasma centrale in Europe

Anaplasma centrale (Rickettsiales: Anaplasmataceae) is used as a live vaccine for cattle against the pathogenic Anaplasma marginale in tropical and subtropical areas. Herein we report a clinical case of bovine anaplasmosis associated with A. centrale infection in Italy, together with the first molecular identification and phylogenetic analysis of this Anaplasma species or subspecies in Europe.

First report of bovine anaplasmosis by Anaplasma centrale in Europe, molecular identification and phylogenetic analysis

First report of bovine anaplasmosis by Anaplasma centrale in Europe, molecular identification and phylogenetic analysis L. Ceci & N. Decaro & E. Lorusso & P. Paradies & G. Elia & V. Martella & C. Buonavoglia & G. Carelli Published online: 12 August 2008 # Springer Science + Business Media B.V. 2008

Pan-Mediterranean comparison for the molecular detection of Theileria annulata.

Abstract: Seven laboratories decided to compare their molecular diagnostic techniques to identify Mediterranean theileriosis caused by Theileria annulata. Each laboratory used either PCR or PCR and reverse line blot hybridization (RLB) to identify T. annulata. Five laboratories sent their own samples to laboratory 4 to be recoded and passed on to at least two other laboratories. A total of 120 blood samples were analyzed during this study, generating 540 results. Laboratory 1 sent only T. annulata‐infected samples (positive control batch), and all the laboratories testing this batch found 100% infection. Laboratory 2 sent only negative samples from a Mediterranean area where T. annulata was unknown, and two laboratories out of three found a few positive samples in these negative samples. For the remaining samples, detection performance was variable. Agreement between laboratories ranged from 21.4 to 91.3%. The overall mismatch between laboratories was around 30% by whatever technique used. This paper describes the methodological parameters that could explain the variation of results.