Comparison of different direct diagnostic methods to identify Babesia bovis and Babesia bigemina in animals vaccinated with live attenuated parasites

Blood smear examination, flow cytometry, duplex Polymerase Chain Reaction (PCR), and duplex nested PCR (nPCR) were evaluated for detection of Babesia bigemina and Babesia bovis infections in cattle vaccinated with live attenuated strains. Two groups of four cattle were immunized with either B. bigemina (Bi) or B. bovis (Bo). On day 23 post inoculation (PI), Bi cattle were vaccinated with B. bovis (BiBo) and Bo cattle were vaccinated with B. bigemina (BoBi). Babesia bigemina was first detected by blood smear examination 7.5+/-3.5 days PI in the Bi group and 32.2+/-1.7 days PI in the BoBi group. The first occurrence of B. bovis in blood smears was 8.0 days PI in the Bo group and 36.0+/-2.6 days PI in the BiBo group. Flow cytometry detected parasitized erythrocytes on day 1.7+/-1.5 and 2.2+/-1.5 PI in the Bi and Bo groups, respectively, but did not discriminate between the two Babesia spp. Duplex PCR detected B. bigemina on day 4.0+/-0.8 and 26.0+/-0.8 PI in the Bi and BoBi groups, respectively, and B. bovis on day 4.0 and 25.3+/-0.5 PI in the Bo and BiBo groups, respectively. The duplex nPCR detected B. bigemina on 3.0+/-0.8 and 25.0+/-0.0 days PI in the Bi and BoBi groups, respectively, and 4.7+/-1.7 and 27.7+/-6.2 days PI in the Bo and BiBo groups, respectively. Duplex nPCR outperformed the other tests in terms of specificity and sensitivity, indicating that it is the most useful method for identifying Babesia spp. in cattle following vaccination.     

Response to Change: Understanding How Forest Management Activities Affect Nonindigenous Species Occurrence and Distribution

ABSTRACT

Disturbances, including natural phenomena and forest management activities such as thinning, reforestation, rehabilitation seeding, and livestock grazing, are often associated with changes that make systems more susceptible to invasion by nonindigenous species (NIS). However, the susceptibility of a landscape to any particular species is not uniform. To prevent and manage (NIS) effectively we need to understand the effect of different types of disturbance on such species and understand how these effects vary on the landscape. Such an understanding is required in a general way for different nonindigenous taxa, but also for specific species or species associations in order to be able to develop comprehensive management plans. We performed three reviews, one each for nonindigenous plants, pathogens, and insects concerning the effect of a range of different forest management activities on their occurrence, spread, and spatial distribution in the United States of America. Much remains to be understood about the processes and patterns of invasion at the finer spatial landscape scales (e.g., < 50 m) that are most relevant to management decisions for a specific area.     

Validation of a fecal glucocorticoid metabolite assay for collared peccaries (Pecari tajacu)

The possibility of assessing endogenous adrenal activity in the collared peccary (Pecari tajacu) was tested by using an adrenocorticotropic hormone (ACTH) challenge in a fecal glucocorticoid metabolite (FGM) assay. Feces were collected from 12 captive adult male peccaries beginning 48 hr prior to challenge; six of these animals received the challenge as an ACTH injection and the other six were injected with saline solution. Feces collection ended 120 hr after injections. As a control, feces were collected for eight consecutive days from another six adult male peccaries that remained in their original mixed-sex herds in semiconfined paddocks. All feces samples were freeze-dried, extracted by an ethanol vortex method, and assayed for glucocorticoids by means of an enzyme immunoassay. FGM concentrations were compared between the treatments by a repeated measures analysis of variance (ANOVA) followed by a post hoc Tukey test. The assay is reliable but, instead of the usual proportion of 1:50 in ethanol (fecal mass:solvent), 1:10 is recommended for best extraction of FGM. Baseline FGM concentrations were similar among the ACTH, saline, and control treatments (29.7 +/- 11.2 ng/g(-1) dry feces) during the 48 hr before the challenge. The ACTH group reached an FGM excretion peak at 24 hr post-treatment, followed by a decline, while in the control and saline groups FGM levels remained relatively constant. Therefore, the fecal glucocorticoid metabolite assay reflects endogenous adrenal activity in the collared peccary and is a powerful tool for noninvasive stress monitoring in peccaries.     

Colour Doppler ultrasound imaging of blood flows variations in neoplastic and non‐neoplastic testicular lesions in dogs

Testicular tumours are the most common neoplasms in male dogs accounting for approximately 90% of all tumours affecting the genitourinary tract. Gray‐scale ultrasonography in combination with colour and power Doppler imaging has been well accepted as an accurate technique for assessing scrotal lesions and vascularization of the testis. Colour Doppler sensitivity for low blood flows appears promising in the study of testicular disorders. The aim of this study was to assess if colour and power Doppler ultrasound is a good tool for the investigation of testicular lesions in dogs, to report the sonographic features of lesions and to measure colour and power Doppler parameters such as resistive index (RI), pulsatility index (PI), hypovascularization and hypervascularization (VI) determining if they can be used to distinguish testicular neoplasms from the wide spectrum of non‐neoplastic pathological findings. In this study, 50 male dogs of various breeds, aged between 7 and 14 years, presented with testicular disorders were selected. RI and PI were calculated. Mean RI values for neoplastic, inflammatory and degenerative lesions were 0.54, 0.45 and 0.58, respectively. Mean PI values were 0.62, 0.55 and 0.63, respectively. Hypovascularization and hypervascularization of the lesion were evaluated throughout the vascularity index (VI). Vascular signals in neoplasms were significantly intensified around and inside the mass if compared with those measured during inflammatory and degenerative lesions. VI markedly increased in solid tumours. Pathological testes were removed; macroscopical, histological and immunoistochemical evaluations were carried out. Colour Doppler showed increased intralesional and peripheral flows in all neoplastic lesions analysed. No flows were detected around cysts.     

Cross-reactivity of mAbs to human CD antigens with sheep leukocytes

A panel of 377 commercially available mAbs was submitted to the animal homologue section of the Eighth International Workshop on Human Leukocyte Differentiation Antigens (HLDA8, Adelaide, Australia) for cross-reactivity studies in a range of vertebrate species. Eight commercial suppliers participated by providing isotype controls and mAbs specific for a total of 144 CD antigens. In this study, we describe the results of flow cytometric testing of the reactivity of these mAbs with leukocyte populations isolated from blood, bronchoalveolar lavage, and ileal Peyer's patches of sheep. A total of 52 mAbs were identified as potentially reacting with sheep blood leukocytes in the first round of screening with blood leukocytes. In the second phase, reactivity of selected mAbs was further analyzed by repeating the screening with blood leukocytes at an independent facility. Screening of selected mAbs for reactivity with myeloid antigens was completed with alveolar macrophages and screening for reactivity with B cell antigens was completed with ileal Peyer's patch B cells. This screening identified mAbs that consistently reacted with both putative myeloid (CD10, CD22, CD23, CD27, CD29, CD32, CD49d, CD81, CD86, CD88, CD163, CD165) and B cell (CD10, CD22, CD23, CD27, CD29, CD32, CD49d, CD81, CD86, CD88, CD165) activation or differentiation antigens. Further studies will be required to determine if each mAb cross-reacts with an orthologous leukocyte antigen.     

Evaluation of antioxidant effect of extracts of Symphyopappus casarettoi

The ethanolic extract of Symphyopappus casarettoi syn. Eupatorium casarettoi was partionated in hexanic, chloroformic and methanolic fractions. Extract and fractions were tested for their antioxidant activity in vitro and ex vivo assays. The methanolic fraction showed a higher antioxidant potential compared to the others fractions, which was correlated with its total phenol content. In addition, the ethanolic extract and the methanolic fraction attenuated ex vivo iron-induced cell death, quantified by lactate dehydrogenase leakage, and effectively protected against lipid damage induced by iron. These findings suggest that the ethanolic extract of S. casarettoi inflorescence and its methanolic fraction have in vitro and ex vivo antioxidant properties.