Anaplasma phagocytophilum in horses and ticks: a preliminary survey of Central Italy

Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, affects several species of wild and domesticated mammals, including horses. In this work we compared direct and indirect methods to evaluate A. phagocytophilum presence in Central Italy: 135 sera were screened by IFA for A. phagocytophilum and other haemopathogens (Theileria equi and Babesia caballi). Each horse was also tested for A. phagocytophilum 16S rRNA with a nested-PCR technique. In order to examine the risk of A. phagocytophilum transmission, 114 ticks were examined for the presence of A. phagocytophilum by PCR targeting the 16S rRNA. The seroprevalence against A. phagocytophilum was 17.03% and 11 horses (8.14%) showed positive PCR results. The concordance rate of A. phagocytophilum detection between IFAT and PCR had a K value of 0.34.     

Molecular detection of Leishmania sp. in cats (Felis catus) from Andradina Municipality, São Paulo State, Brazil

The aim of this work was to molecularly detect Leishmania species in 52 cats from Andradina Municipality, S?o Paulo State, Brazil. The direct parasitological test was performed by using imprints of poplited lymph node, bone marrow and spleen to verify amastigote forms of Leishmania spp. The samples that were positive parasitological tests were subjected to molecular analysis (PCR) and sequencing. Infection was detected for 5.76% (3/52) of the examined cats and two had presence of amastigote forms of Leishmania spp. in lymph nodes. Polymerase chain reaction (PCR) of kinetoplast minicircle DNA, indicated positive amplification for samples of spleen and lymph nodes and the sequencing resulted in 97% similarity with Leishmania (L.) chagasi. This study proved the occurrence of infection with Leishmania (L.) chagasi in felines from Andradina municipality, S?o Paulo State.     

In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M –phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.     

Response to domestication and selection for growth in the European sea bass (Dicentrarchus labrax) in separate and mixed tanks

Selective breeding of European sea bass (Dicentrarchus labrax) receives a growing interest, as the estimated heritability of growth is medium to high. In this study, we compared the offspring of four groups of sea bass sires, mated with the same wild dams: wild (W), first generation of domestication (D), first generation of mass selection for length (M), first generation of PROSPER-like selection for length (P). The comparison was done both in replicated tanks (separate rearing) and in mixed tanks (mixed rearing) where sire origins were recovered by genotyping of eight microsatellite markers. Weight, length and growth rate were measured from day 238 post-fertilization (69 g mean weight) to day 611 post-fertilization (390 g mean weight). Both in mixed and separate tanks, both selected groups (P, M) were larger than unselected groups (W, D). No difference was seen at any time between W and D, nor between M and P. The selection response estimate on weight was larger in mixed tanks when compared to separate tanks (+ 42% in mixed tanks, + 23% in separate tanks at day 611), yielding realized heritability estimates of 0.60 and 0.34, respectively, and confirming the excellent potential of the species for growth improvement through selective breeding. Both selection response and the amplification effect between mixed and separate tanks decreased as rearing density increased. Our hypothesis is that selection response is magnified by competition in mixed tanks, while sub-optimal rearing conditions lower the observed selection response, more in separate tanks (where selected thus larger fish are at a higher density than unselected ones) than in mixed tanks (where all fish experience the same density effects).     

Comparison of histologic techniques for the diagnosis of bovine tuberculosis in the framework of eradication programs.

Rapid diagnosis of tuberculosis in cattle reacting positive in antemortem assays is crucial in countries where eradication programs are operated to confirm the presence of the infection in tuberculosis-free herds. This study evaluated the accuracy of histopathologic examination by hematoxylin and eosin and Ziehl-Neelsen (ZN) staining applied in this framework, when suspected lesions are caused by low infectious doses and are detected in early stages of the disease. For this purpose, histologic methods were compared with mycobacterial culture as reference test on suspected lymph node samples from 173 cattle reacting positive in antemortem tests. Histopathology demonstrated high sensitivity (93.4%) and specificity (92.3%), while ZN sensitivity and specificity were respectively 33.9% and 100%. There was good agreement between histopathology and bacterial culture, suggesting that histopathologic examination is a reliable tool for rapid diagnosis in countries where active tuberculosis eradication programs allow the prompt identification and elimination of reactor cattle. Histopathology permits identification of typical mycobacterial lesions and its differentiation from other causes.     

Avian influenza epidemiology in semi-intensive free ranging duck flocks of the Moyingyi Wetland in Bago East District,Myanmar

Highly pathogenic avian influenza virus subtype H5N1 first entered Myanmar in 2006 in the Mandalay District. Several H5N1 outbreaks followed and the one of Bago East District (2007) required post outbreak surveillance in the at-risk domestic duck population of the Moyingyi Wetland. A field epidemiological study based on a randomised prospective stratified study with five surveys provided the serological evidence that the avian influenza H5 subtype circulates in the domestic duck population and spreads to almost all the newly housed (and negative) flocks in the time span of a seasonal production cycle. Virological investigation was negative. The survival analysis showed that the probability of seroconversion increased rapidly over the study period, without significant difference among different agro-ecosystems. The analysis suggests that viral spread in the new cycle could be limited if control measures were adopted at the time new flocks are housed. The study recommends that future surveillance schemes for ducks are designed in a way to get as much information as possible from serological results which should drive virological sampling to determined farms.     

Recent spreading of a divergent canine parvovirus type 2a (CPV-2a) strain in a CPV-2c homogenous population

Canine parvovirus type 2 (CPV-2), which causes acute hemorrhagic enteritis in dogs, is comprised of three antigenic variants (2a, 2b, and 2c) that are distributed worldwide with different frequencies. Variant prevalence was analyzed in 150 CPV-2-positive samples collected from the Uruguayan dog population in 2007-2010. Samples were analyzed with polymerase chain reaction, restriction fragment length polymorphism, and sequencing of the coding region for the largest and most variable loop of the VP2 capsid protein. CPV-2c was the only strain detected from 2007 to 2009. Uruguayan CPV-2c showed high homogeneity in both nucleotide and amino acid sequences, indicating a low level of genetic variability. In 2010, an unexpected epidemiological change occurred in Uruguay as a consequence of the appearance of a novel CPV-2a strain. This variant rapidly spread through the Uruguayan dog population and was detected in 20 of the 52 cases (38%) analyzed in 2010. CPV-2a sequences were identical in all field viruses analyzed, and in addition to the characteristic 426Asn residue, the sequences showed amino acid substitutions (267Tyr, 324Ile, and 440Ala) not observed in the Uruguayan CPV-2c. These data and the first detection in April 2010 suggest that the CPV-2a variant recently emerged in Uruguay and underwent clonal expansion. This observation is the first case in which a CPV-2a variant increased its frequency in a dog population where CPV-2c was prevalent. Our results emphasize the dynamic changes in CPV variants and highlight the importance of ongoing surveillance programs to provide a better understanding of virus epidemiology.     

Intra-abdominal leiomyosarcoma in a ferret (Mustela putorius Furo): histopathological and immunohistochemical characterization

A 5 years old female ferret with an abdominal palpable mass confirmed at echo-graphic examination died during an explorative laparotomy. A single lymph-node-like nodule was found adjacent to the intestinal loops. The round mass well circumscribed, solid and white, histologically, at low magnification, appeared encapsulated and built up by a population of atypical spindle cells arranged in interwoven fascicles. The cells had high anisocytosis degree, moderate mitotic activity and prominent nucleoli. A central area of necrosis was present. To characterise the tumour immunohistochemically cytokeratin, vimentin, S-100, melan-A, vWF, desmin, actin and alpha-actin were applied. Neoplastic cells resulted positive to vimentin, actin and alpha-actin. Based on the histological and immunohistological pattern a diagnosis of leiomyosarcoma was made.