Immune responses of immunocompetent and immunocompromised mice experimentally infected with Bartonella henselae

The aim of this study is to understand host immune responses in immunocompetent and immunocompromised mice against Bartonella henselae infection. BALB/c and nude (BALB/c nu/nu) mice were inoculated intraperitoneally with 10(8) colony forming units of B. henselae (Houston-1 strain). Blood, brain, liver, spleen, kidney and bone marrow samples were collected 0, 3, 7, 14, 21 and 28 days after infection and submitted to bacteriological, serological and genetical examinations. B. henselae was isolated only from the liver 3 days after infection. DNA of the inoculums was detected by polymerase chain reaction from blood, liver, and spleen samples collected from BALB/c and blood from nude mice 3 and 7 days after infection. No bacterial DNA was detected from both BALB/c and nude mice thereafter during 4 weeks observation periods. These results indicate that the T-cell may not participate in the effective elimination of the organisms from mice. In addition, western blot analysis revealed that the antigens of 27.3- and 31.5-kDa reacted with IgM antibodies from the blood of BALB/c and nude mice after 3 days of infection, suggesting that these antigens were recognized by thymus-independent mechanism. Furthermore the antigens were detected from the culture-supernatants of B. henselae, indicating that these antigens were secreted from the organisms.     

Serological evidence of infection of Anaplasma and Ehrlichia in domestic animals in Xinjiang Uygur Autonomous Region area, China

Serological methods were utilized to detect Anaplasma and Ehrlichia infection in domestic animals in Xinjiang Uygur Autonomous Region, China. By using an indirect immunofluorescence assay (IFA), antibodies that reacted with Anaplasmaphagocytophilum and Ehrlichiachaffeensis were detected mainly in ruminants kept on pastureland in Altai, Ili and Kashgar area. Antibody titers up to 1:320 were recorded. These results indicate that ruminants kept in these areas may be infected with some species of Anaplasma and Ehrlichia.     

Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50

The surface antigen P50 of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding truncated P50 (P50t) lacking a signal peptide and C-terminal hydrophobic regions were cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). More than 90% portion of the GST-P50t was expressed as a soluble form, in contrast with GST-P50f (full-length), which was completely expressed as an insoluble form. This result indicates that removal of the hydrophobic signal peptide and C-terminus had dramatically improved its hydrophilicity. The purified GST-P50t was tested in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to B. gibsoni in dogs. The ELISA with GST-P50t clearly differentiated between B. gibsoni-infected dog sera and uninfected dog sera. In addition, the ELISA detected no cross-reactivity with sera from dogs experimentally infected with the closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi. Field serum samples collected from dogs in Japan and China were examined for the diagnosis of B. gibsoni infection by using the ELISA. 14.5% (9/62), 5.8% (7/120), and 5.4% (2/37) of tested samples were positive for dogs from Okinawa, Yamaguchi, and Osaka prefectures, Japan, respectively. On the other hand, 4.8% (2/41) of tested samples were positive for dogs from Nanjing, China. These results suggest that the GST-P50t could be a reliable reagent for practical use in ELISA for the serodiagnosis of canine piroplasmosis caused by B. gibsoni.     

Differences in flowering phenology and seed production of four morning glory (Ipomoea) species in Japan

The spread of morning glory (Ipomoea spp.) in soybean fields in Japan has severely decreased soybean yield. Yet, current control measures do not control the proliferation of Ipomoea spp. As little is known about the flowering period and seed production among the different invading Ipomoea spp., it is challenging to create targeted control measures based on ecological characteristics. This study aimed to reveal the characteristics of the flowering phenology and seed production of four morning glory species, namely, Ipomoea coccinea L. (red morning glory), Ipomoea lacunosa L. (pitted morning glory), Ipomoea hederacea L. Jacq. var. integriuscula A. Gray (entireleaf morning glory), and Ipomoea triloba L. (three-lobe morning glory). Between 2017 and 2019, the four selected study species were grown under similar conditions of soil quality, irrigation, and environmental influences and their flowering phenology and seed data were recorded. The flowering period ranged from 36 to 40 days, and the initial flowering of I. triloba was approximately 2 weeks later than the others. I. coccinea had the highest flowering number and seed production, followed by I. lacunosa, I. triloba, and I. hederacea var. integriuscula. The fruit setting rate of I. triloba decreased later in the reproductive stage but tended to increase as the daily mean temperature increased on each flowering day. Thus, we report that the flowering phenology and seed production differed greatly among the Ipomoea spp. These findings can provide crucial insights into designing targeted species-specific control measures against the spread of Ipomoea spp. in Japan.     

Seroprevalence of Toxoplasma gondii antibodies in stray cats and dogs in the Bangkok metropolitan area, Thailand

Cats and dogs are the most popular pet animals worldwide. Cats are the natural reservoir of Toxoplasma gondii and excrete the resistant oocyst to environments. On the other hands, dogs play a role in the mechanical transmission of the parasite. Stray cats and dogs in the Bangkok metropolitan area are becoming a public concern because there is a considerable increase in their number annually. These facts indicate the risk of mechanically spreading zoonoses including toxoplasmosis to humans since human acquire the infection from infected mammals, either directly or indirectly. In the present study, the presence of T. gondii antibodies was examined in 592 cats and 427 dogs from October 2001 to September 2002 by using a latex agglutination test. T. gondii antibodies were detected in 65 (11.0%) of the 592 cats and 40 (9.4%) of the 427 dogs. The antibody titers in the positive animals ranged from 1:64 to 1:2048. Seroprevalence was significantly higher in female cats than in male cats. The present study suggested that T. gondii was widespread in the stray animals in the Bangkok metropolitan area; therefore, it is essential to control the number of stray cats and dogs in order to reduce the transmission of toxoplasmosis to animals and humans.     

Serological survey of Ehrlichia and Anaplasma infection of feral raccoons (Procyon lotor) in Kanagawa Prefecture, Japan

Numbers of feral raccoon; the possible reservoir animal of Ehrlichia and Anaplasma, are increasing in Japan. Thus serological methods were utilized to examine Ehrlichia and Anaplasma infection in raccoons from Kanagawa Prefecture, Japan. By using an indirect immunofluorescence assay, among 187 feral raccoons examined, 1 (0.5%) serologically reacted with Ehrlichia canis, 3 (1.6%) with Ehrlichia chaffeensis and 1 (0.5%) with Anaplasma phagocytophilum with the titers of 1:40 or more. Although screening PCR for Ehrlichia and Anaplasma species failed to detect the presence of ehrlichial DNA in serum samples, results of the serological tests suggested that the feral raccoons might be infected with some species of Ehrlichia and Anaplasma.     

Persorption mechanisms of luminal antigenic particulates via apoptotic epithelial cells of intestinal villi into systemic blood circulation in orally immunized rats

The possibility of persorption of prefixed bovine serum albumin-coated sheep erythrocytes (BSA-SEs) from mucous epithelial cells and its mechanisms were investigated in rats orally immunized by BSA for 14 consecutive days. On the day after the final oral immunization, the rats were duodenally perfused by BSA-SEs or non-coated SEs. BSA-SEs were also duodenally perfused in non-immunized rats. Thirty min after perfusion, BSA-SEs were significantly more engulfed by late-apoptotic-stage villous columnar epithelial cells in the orally immunized rats than those in other experiments. The specific antibody (SpAb) was detected on the surfaces of BSA-SEs in rats with oral immunization. In Peyer's patches of all animals, no SEs reached the follicle-associated epithelium, because of the close attachment of follicle-associated intestinal villi and the thick mucous layer. BSA-SEs were more frequently persorbed into portal blood in the orally immunized rats than in other rats. Small numbers of BSA-SEs or SEs were detected in the systemic blood of all animals. BSA-SEs were also histologically found in the blood vessels of the liver, but not in mesenteric lymph nodes. These findings suggest that sensitized antigenic particulates are taken up by late-apoptotic-stage villous columnar epithelial cells in the small intestine and are finally persorbed into the systemic blood circulation. The uptake of antigenic particulates might be mediated by its luminal SpAb.     

Characterization of the suppressive effects of the biological control strain VAR03-1 of <Emphasis Type="Italic">Rhizobium vitis</Emphasis> on the virulence of tumorigenic <Emphasis Type="Italic">R. vitis</Emphasis>

Rhizobium vitis: strain VAR03-1 is a biological control agent that suppresses grapevine crown gall disease caused by a tumorigenic strain of R. vitis (Ti). Both acetosyringone-induced expression of a virulence gene and the growth of Ti were suppressed in vitro when it was cultivated in the VAR03-1 culture filtrate. These inhibitory effects were reduced by high-temperature treatment or incubation for 72 h. Both activities were detected in the high molecular weight fraction (>?100 kDa) of the filtrate. Our results suggest that the antagonistic effects of VAR03-1 on Ti are mediated by large particle(s) released in the culture media.     

The influence of fig proteases on the inhibition of angiotensin I‐converting and GABA formation in meat

The purposes of this research were to use fig protease for texture tenderizing, and to inhibit angiotensin I‐converting enzyme (ACE) action and γ‐aminobutyric acid (GABA) formation of meat. Liberated peptides by the enzymatic action of fig protease in processing meat and free amino acids were determined and ACE inhibitory activity was assayed. Meat treated with fig protease became tender as indicated by shear force value (SFV) which was half of those of non‐fig treated meat during storage even at 5°C. Liberated peptides, free amino acids and GABA increased while extremely low levels of Glu were detected after storage. The optimal temperature of fig protease against meat was 80°C. However, the activity of fig protease decreased after pre‐heating more than 40°C. High ACE inhibitory activity of a mixture of fig and meat was found around 80°C, and the value corresponded to the amount of liberated peptide. A lot of liberated peptides were found at 60–80°C and pasterization of meat product becomes convenient to produce peptides. Production of ACE inhibitory peptides and GABA can be expected as the healthy functional meat product such as antihypertensive activity and improve brain function.     

A serine proteinase from the sarcoplasmic fraction of red sea bream Pagrus major is possibly derived from blood

Collagen degradation is known to be involved in the post mortem tenderization of fish muscle. A serine proteinase that is assumed to be related to collagen degradation after fish death was purified from the sarcoplasmic fraction of red sea bream Pagrus major by ammonium sulfate fractionation and column chromatography on Sephacryl S-300, Q Sepharose and Phenyl Sepharose CL-4B. The enzyme hydrolyzed gelatin and was obtained as a protein band of approximately 38 kDa upon sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions. The N-terminal amino acid sequence of the enzyme was determined for 32 residues. A protein that had the same N-terminal amino acid sequence as the enzyme for ten residues was purified from the serum of red sea bream and showed the same characteristics as the enzyme. Therefore, it is suggested that the serine proteinase migrates from the blood to muscle and degrades muscle proteins after the death of the fish.