Molecular epidemiological study of gammaherpesvirus in domestic cats in Japan.

Gammaherpesviruses (GHVs) are members of an emerging subfamily of the family Herpesviridae. A recent study identified a novel GHV in domestic cats (Felis catus GHV1, FcaGHV1), and epidemiological surveys have found that FcaGHV1 is distributed worldwide. In this study, we investigated the prevalence of GHVs in domestic cats in Japan with a molecular epidemiological survey. Blood samples were collected from 1,738 domestic cats and GHV-derived DNA was detected with PCR in 1.3% (23/1,738) of the Japanese domestic cats. The FcaGHV1 detected in this study was very similar to FcaGHV1 detected in a domestic cat in North America. Older age (>5 years old) and Feline immunodeficiency virus infection were identified as risk factors for GHV infection.

Detection and molecular characterization of Babesia, Theileria, and Hepatozoon species in hard ticks collected from Kagoshima, the southern region in Japan.

To reveal the distribution of tick-borne parasites, we established a novel nested polymerase chain reaction (PCR) system to detect the most common agents of tick-borne parasitic diseases, namely Babesia, Theileria, and Hepatozoon parasites. We collected host-seeking or animal-feeding ticks in Kagoshima Prefecture, the southernmost region of Kyusyu Island in southwestern Japan. Twenty of the total of 776 tick samples displayed a specific band of the appropriate size (approximately 1.4-1.6kbp) for the 18S rRNA genes in the novel nested PCR (20/776: 2.58%). These PCR products have individual sequences of Babesia spp. (from 8 ticks), Theileria spp. (from 9 ticks: one tick sample including at least two Theileria spp. sequences), and Hepatozoon spp. (from 3 ticks). Phylogenetic analyses revealed that these sequences were close to those of undescribed Babesia spp. detected in feral raccoons in Japan (5 sequences; 3 sequences being identical), Babesia gibsoni-like parasites detected in pigs in China (3 sequences; all sequences being identical), Theileria spp. detected in sika deer in Japan and China (10 sequences; 2 sequences being identical), Hepatozoon canis (one sequence), and Hepatozoon spp. detected in Japanese martens in Japan (two sequences). In summary, we showed that various tick-borne parasites exist in Kagoshima, the southern region in Japan by using the novel nested PCR system. These including undescribed species such as Babesia gibsoni-like parasites previously detected in pigs in China. Importantly, our results revealed new combinations of ticks and protozoan parasites in southern Japan. The results of this study will aid in the recognition of potential parasitic animal diseases caused by tick-borne parasites.

Hypoxia-activated prodrug TH-302 decreased survival rate of canine lymphoma cells under hypoxic condition.

We tested the hypotheses that hypoxic stimulation enhances growth potentials of canine lymphoma cells by activating hypoxia-inducible factor 1α (HIF-1α), and that the hypoxia-activated prodrug (TH-302) inhibits growth potentials in the cells. We investigated how hypoxic culture affects the growth rate, chemoresistance, and invasiveness of canine lymphoma cells and doxorubicin (DOX)-resistant lymphoma cells, and influences of TH-302 on survival rate of the cells under hypoxic conditions. Our results demonstrated that hypoxic culture upregulated the expression of HIF-1α and its target genes, including ATP-binding cassette transporter B1 (ABCB1), ATP-binding cassette transporter G2 (ABCG2), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), and survivin, and enhanced the growth rate, DOX resistance, and invasiveness of the cells. Additionally, TH-302 decreased the survival rate of the cells under hypoxic condition. Our studies suggest that hypoxic stimulation may advance the tumorigenicity of canine lymphoma cells, favoring malignant transformation. Therefore, the data presented may contribute to the development of TH-302-based hypoxia-targeting therapies for canine lymphoma.

The presence of tick-borne diseases in domestic dogs and cats living on Iriomote-jima and Tsushima islands.

The Iriomote cat and Tsushima leopard cat are endangered wildcats in Japan and inhabit only Iriomote-jima and Tsushima islands, respectively. Domestic dogs and cats living on Iriomote-jima and Tsushima islands were surveyed to clarify the interrelationship between wildcats and domestic animals regarding tick-borne disease transmission. Pathogen-derived DNA in blood samples was detected by polymerase chain reaction. Babesia gibsoni was detected in dogs of Iriomote-jima, and Hepatozoon felis and hemoplasmas were detected in domestic cats of Tsushima. Because the H. felis detected in this study was closely related to that isolated from wildcats, we suspect that common H. felis is harbored and transmitted among wildcats and domestic cats via ticks in Tsushima.

A molecular epidemiological survey of Babesia, Hepatozoon, Ehrlichia and Anaplasma infections of dogs in Japan.

Tick-borne diseases are often encountered in canine clinical practice. In the present study, a molecular epidemiological survey of dogs in Japan was conducted to understand the prevalence and geographical distribution of Babesia spp., Hepatozoon spp., Ehrlichia spp. and Anaplasma spp. Pathogen-derived DNA in blood samples obtained from 722 dogs with a history of exposure to ticks and/or fleas was examined by PCR. The prevalence of Babesia gibsoni, Babesia odocoilei-like species, Hepatozoon canis and Ehrlichia spp./Anaplasma spp. was 2.4% (16/722), 0.1% (1/722), 2.5% (18/722) and 1.5% (11/722), respectively. While B. gibsoni and Ehrlichia spp./Anaplasma spp. were detected in the western part of Japan, H. canis was detected in Tohoku area in addition to western and central parts of Japan.

Molecular survey of arthropod-borne pathogens in ticks obtained from Japanese wildcats.

The Iriomote cat (IC), Prionailurus bengalensis iriomotensis, and the Tsushima leopard cat (TLC), Prionailurus bengalensis euptilurus, are endangered subspecies of leopard cats in Japan. In addition to habitat destruction and road kills, infectious diseases may threaten their populations, and infection with arthropod-borne pathogens has been reported in both subspecies. Infestations with ectoparasites, especially ticks, have frequently been observed in ICs and TLCs. In the present study, ticks collected from captured ICs and TLCs between November 2011 and January 2012 were morphologically identified and the prevalence of the pathogens Bartonella sp., Babesia sp., Ehrlichia sp., Anaplasma sp., Hepatozoon sp., and hemoplasmas in the ticks was molecularly evaluated. The ticks Haemaphysalis longicornis, H. hystricis, and Amblyomma testudinarium were obtained from ICs, and H. megaspinosa, Ixodes tanuki, H. campanulata, and A. testudinarium were collected from TLCs. The pathogens Hepatozoon felis, Babesia sp., and Anaplasma bovis were detected in ticks obtained from ICs, while H. felis. Babesia sp., Ehrlichia sp., E. muris, 'Candidatus Mycoplasma haemominutum', and Bartonella henselae were found in ticks from TLCs. To protect and conserve these endangered animals, continuous monitoring and additional surveys will be necessary to understand the role of ticks as disease vectors in Japanese wildcats.

The clinical utility of two human portable blood glucose meters in canine and feline practice.

BACKGROUND: Portable blood glucose meters (PBGMs) are useful for serial measurements of blood glucose and creation of blood glucose curves in veterinary practice. However, it is necessary to validate PBGMs designed for people for veterinary use. OBJECTIVES: Our objective was to evaluate the accuracy of 2 PBGMs designed for people for use in dogs and cats. METHODS: The blood glucose levels were determined in blood samples collected from 69 dogs and 26 cats admitted to the Kagoshima University Veterinary Teaching Hospital, using a MEDISAFE [PBGM-T] and an Antsense III [PBGM-H], and a FUJI DRI-CHEM 7000V as reference method. The correlations and agreements among the results were statistically analyzed. RESULTS: Simple regression analyses revealed a high correlation between values from both PBGMs and the reference method in both dogs and cats. However, Passing-Bablok regression and Bland-Altman analyses revealed that the data from both PBGMs did not show statistical agreement with the reference values. Concordance correlated coefficients were moderate for the PBGM-T and almost perfect for the PBGM-H for canine samples, and were poor for the PBGM-T and substantial for the PBGM-H for feline samples. Hematocrit values significantly affected the results of the PBGM-T, but not the PBGM-H. Error grid analyses revealed that all measurements from both PBGMs would lead to acceptable treatment decisions. CONCLUSIONS: Our findings suggest that both PBGMs, especially the PBGM-H, would be clinically useful in small animal practice, although there was a bias between each PBGM and the reference method.

Molecular epidemiologic survey of Bartonella, Ehrlichia, and Anaplasma infections in Japanese Iriomote and Tsushima leopard cats.

The Iriomote cat (IC; Prionailurus iriomotensis) and the Tsushima leopard cat (TLC; Prionailurus bengalensis euptilura) are endangered wild felids in Japan. As a part of ongoing conservation activities, we conducted a molecular, epidemiologic survey of Bartonella, Ehrlichia, and Anaplasma infections in wild IC and TLC populations. Blood samples (47 from 33 individual IC; 22 from 13 TLC) were collected between August 2002 and January 2011. Using PCR analysis, we confirmed the presence of Bartonella henselae in ICs and Bartonella clarridgeiae in TLCs, with prevalences of 6% and 8%, respectively. Using PCR and basic local alignment search tool analyses, we identified Ehrlichia canis in both cats and Anaplasma bovis in TLCs. The prevalence of E. canis was 12% in ICs and 8% in TLCs, and the prevalence of A. bovis was 15% in TLCs. This is the first report, to our knowledge, of B. henselae, B. clarridgeiae, E. canis, and A. bovis infections in these two endangered species. Continuous monitoring of these pathogens is needed for their conservation.

Epidemiological survey of tick-borne protozoal infection in iriomote cats and tsushima leopard cats in Japan.

This epidemiological survey was conducted to determine the prevalence of Hepatozoon, Babesia and Theileria infection in the Iriomote cat (IC) and the Tsushima leopard cat (TLC). Blood samples from 43 ICs and 14 TLCs were collected between November 2002 and January 2012. Polymerase chain reaction and DNA sequencing analyses detected a Hepatozoon felis infection prevalence of 72.0% (31/43 cats) and 100% (14/14 cats) in ICs and TLCs, respectively. The degree of Hepatozoon parasitemia observed on blood smears ranged from 0.1 to 4.7%. However, no cases had obvious clinical signs of hepatozoonosis. Neither Babesia- nor Theileria-infected wildcats were detected in this study.

An epidemiological survey of hemoplasma infection in Iriomote cats (Prionailurus bengalensis iriomotensis).

An epidemiological survey of Iriomote cats (Prionailurus bengalensis iriomotensis) was conducted to understand the prevalence and molecular characteristics of hemotropic mycoplasma (hemoplasma). A series of ecological surveys of Iriomote cats were performed between November 2003 and September 2010. During this period, 31 Iriomote cats were captured or found, and 39 blood samples were collected. Polymerase chain reaction screening for hemoplasmas and BLAST searches revealed that 4 of the 31 cats were positive for hemoplasma infection (n=3, Mycoplasma haemofelis [Mhf]; n=1, 'Candidatus M. turicensis' [CMt]). The 4 infected cats were captured or found in the northern area of the island of Iriomote. Phylogenetic analyses revealed close relationships between Mhf and CMt isolated from Iriomote cats compared with those from domestic cats and other wild felids. In our study, we identified two species of hemoplasma in Iriomote cats. The number and location of the hemoplasma-positive cats appeared to be limited; however, continuous surveillance of hemoplasma infection in Iriomote cats is necessary.