Journal: BMC Veterinary Research

Article Title: Vector-borne and zoonotic diseases of dogs in North-west New South Wales and the Northern Territory, Australia

doi: 10.1186/s12917-017-1169-2

Figure Lengend Snippet: Exposure variables against microbial result. Frequency table representing the variables of gender, sterilisation status, breed type, age, body condition, PCV and platelet count against Anaplasma platys, Babesia vogeli, Candidatus M. haematoparvum and Mycoplasma haemocanis qPCR positive results, and seropositivity to C. burnetii, Bartonella spp. and Brucella spp.

Article Snippet: Following negative results for commercial PCR testing for Bartonella spp. (IDEXX laboratories, Sacramento), 40 additional fresh blood samples sourced from Yuelamu, Laramba and Alyuen collected from dogs in August 2014 were shipped to the Intracellular Pathogens Research Laboratory, College of Veterinary Medicine, North Carolina State University, USA for Bartonella testing.

Techniques:

Journal: Pathogens

Article Title: Bartonella henselae Detected in Malignant Melanoma, a Preliminary Study

doi: 10.3390/pathogens10030326

Figure Lengend Snippet: Bartonella spp. serology and Bartonella alpha proteobacteria enrichment blood culture PCR results for eight melanoma patients from Oregon.

Article Snippet: : nuclear dye DAPI, anti- Bartonella henselae made in mouse at 1:100 (AB704, Abcam Cambridge, MA, USA) plus anti-mouse IgG Alexa 488 made in donkey at 1:400 dilution (Jackson ImmunoResearch), anti-VEGFC made in goat at 1:200 (AB18883 Abcam, Cambridge, MA, USA), plus anti-goat IgG made in donkey conjugated to Alexa 568 at 1:800 (Jackson ImmunoResearch), and anti-LYVE1 made in rabbit at 1:400 (485214 US Biological, Salem, MA, USA), plus anti-rabbit IgG made in donkey conjugated to Alexa 647 at 1:800. and : anti- Bartonella spp. made in chicken at 1:100 dilution (produced by Aves Labs, Inc., Davis, CA, USA), plus anti-chicken IgY conjugated to Alexa 568 at 1:400 dilution (Jackson ImmunoResearch), and nuclear dye TOPRO3 at 1:5000 (T3605, Thermo Fisher, Waltham, MA, USA).

Techniques:

Journal: Pathogens

Article Title: Bartonella henselae Detected in Malignant Melanoma, a Preliminary Study

doi: 10.3390/pathogens10030326

Figure Lengend Snippet: Bartonella henselae was detected in a melanoma biopsy, multi-stained with antibodies to Bartonella henselae , vascular endothelial growth factor C (VEGFC), lymphatic vessel biomarker (LYVE1), and DAPI nuclear dye. A thick tissue section, 80 microns, was multi-stained with ( a ) nuclear stain DAPI (white), ( b ) B. henselae antibodies (green), ( c ) VEGFC antibodies (red), and ( d ) lymphatic vessel biomarker LYVE1 antibodies (magenta) and imaged with laser scanning confocal microscopy. Merged channels, (panels ( e – g )) reveal that immunoreactive- B. henselae (green in ( b , e – g )) co-localizes with the nuclear dye DAPI (white in a , e ), colocalizes with immunoreactive VEGFC (red in ( c , f )) and appears to be inside the lymphatic vessels (magenta in ( d , g )). Sequential tissue sections that received no B. henselae antibodies show no contribution to the fluorescence signal (data not shown). The 40X UPLFLN NA:1.3, 33 microns total Z-stack thickness, 61 steps, 0.55 micron steps. Scale bar = 50 microns.

Article Snippet: : nuclear dye DAPI, anti- Bartonella henselae made in mouse at 1:100 (AB704, Abcam Cambridge, MA, USA) plus anti-mouse IgG Alexa 488 made in donkey at 1:400 dilution (Jackson ImmunoResearch), anti-VEGFC made in goat at 1:200 (AB18883 Abcam, Cambridge, MA, USA), plus anti-goat IgG made in donkey conjugated to Alexa 568 at 1:800 (Jackson ImmunoResearch), and anti-LYVE1 made in rabbit at 1:400 (485214 US Biological, Salem, MA, USA), plus anti-rabbit IgG made in donkey conjugated to Alexa 647 at 1:800. and : anti- Bartonella spp. made in chicken at 1:100 dilution (produced by Aves Labs, Inc., Davis, CA, USA), plus anti-chicken IgY conjugated to Alexa 568 at 1:400 dilution (Jackson ImmunoResearch), and nuclear dye TOPRO3 at 1:5000 (T3605, Thermo Fisher, Waltham, MA, USA).

Techniques: Staining, Biomarker Discovery, Confocal Microscopy, Fluorescence

Journal: Pathogens

Article Title: Bartonella henselae Detected in Malignant Melanoma, a Preliminary Study

doi: 10.3390/pathogens10030326

Figure Lengend Snippet: Melanoma A375 cells co-cultured with B. henselae . ( a ) Melanoma A375 cells were cultured without and ( b ) with B. henselae organisms (green). After 24 h, cultured cells were fixed and immunostained with anti- Bartonella spp. (green). The length of dendritic processes of the Melanoma A375 cells were shorter when cultured with B. henselae (t 2 = 4.18, p < 0.001 ( n = 10). ( c ) At 24 h, immuno-reactive B. henselae (green) appeared to be within as well as outside of melanoma A375 cells. Z-projection of merged Bartonella spp.-immunoreactivity (green), DNA stain TOPRO3 (red), and differential interference contrast (DIC) imaging. Single optical sections were captured with confocal microscopy at 10× UPLSAPO NA: 0.40. Scale bar = 50 microns.

Article Snippet: : nuclear dye DAPI, anti- Bartonella henselae made in mouse at 1:100 (AB704, Abcam Cambridge, MA, USA) plus anti-mouse IgG Alexa 488 made in donkey at 1:400 dilution (Jackson ImmunoResearch), anti-VEGFC made in goat at 1:200 (AB18883 Abcam, Cambridge, MA, USA), plus anti-goat IgG made in donkey conjugated to Alexa 568 at 1:800 (Jackson ImmunoResearch), and anti-LYVE1 made in rabbit at 1:400 (485214 US Biological, Salem, MA, USA), plus anti-rabbit IgG made in donkey conjugated to Alexa 647 at 1:800. and : anti- Bartonella spp. made in chicken at 1:100 dilution (produced by Aves Labs, Inc., Davis, CA, USA), plus anti-chicken IgY conjugated to Alexa 568 at 1:400 dilution (Jackson ImmunoResearch), and nuclear dye TOPRO3 at 1:5000 (T3605, Thermo Fisher, Waltham, MA, USA).

Techniques: Cell Culture, Staining, Imaging, Confocal Microscopy

Journal: Pathogens

Article Title: Bartonella henselae Detected in Malignant Melanoma, a Preliminary Study

doi: 10.3390/pathogens10030326

Figure Lengend Snippet: Bartonella henselae triggers cytokine release during co-culture with melanoma cells. ( a ) An ELISA documented a significant increase in VEGFC expression at 24 h** after inoculation of melanoma cells with B. henselae (Student’s t -test: t 2 = 7, p < 0.01, n = 6). Significance was determined by Student’s t -test. ( b ) VEGFC expression was greater in co-cultures and the increase in VEGFC-expression occurred earlier, namely 24 h, when B. henselae was present as compared to melanoma cells alone. The rate of VEGFC expression when B. henselae was present was significantly greater than that for melanoma cells alone at 24 h (t 4 = 7, p < 0.05), but did not differ from that of melanoma alone after 48 (t 4 = 0.71, p = 0.54) and 72 h (t 4 = 0.25, p = 0.82). n = 6. ( c ) After inoculation of A375 melanoma cells with B. henselae , media was collected and analyzed for interleukin 8 (IL-8) expression at 24, 48, and 72 h. IL-8 expression was significantly increased in response to the presence of Bartonella at 24 and 48 h time points ** (Student’s t -tests: t 4 = 27.08, p < 0.001, and t 4 = 14.84, p < 0.001 for 24 and 48 h, respectively). IL-8 expression was higher in the presence of Bartonella at 72 h, but the difference was not significant (t 4 = 1.72, p = 0.16). n = 6.

Article Snippet: : nuclear dye DAPI, anti- Bartonella henselae made in mouse at 1:100 (AB704, Abcam Cambridge, MA, USA) plus anti-mouse IgG Alexa 488 made in donkey at 1:400 dilution (Jackson ImmunoResearch), anti-VEGFC made in goat at 1:200 (AB18883 Abcam, Cambridge, MA, USA), plus anti-goat IgG made in donkey conjugated to Alexa 568 at 1:800 (Jackson ImmunoResearch), and anti-LYVE1 made in rabbit at 1:400 (485214 US Biological, Salem, MA, USA), plus anti-rabbit IgG made in donkey conjugated to Alexa 647 at 1:800. and : anti- Bartonella spp. made in chicken at 1:100 dilution (produced by Aves Labs, Inc., Davis, CA, USA), plus anti-chicken IgY conjugated to Alexa 568 at 1:400 dilution (Jackson ImmunoResearch), and nuclear dye TOPRO3 at 1:5000 (T3605, Thermo Fisher, Waltham, MA, USA).

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing