bluuebunny/crossref_metadata_2025 huggingface dataset:
Swine influenza: clinical, serological, pathological, and virological cross‐sectional studies in nine farms in Argentina
Authors: Dibárbora Marina , Cappuccio Javier , Olivera Valeria , Quiroga Maria , Machuca Mariana , Perfumo Carlos , Pérez Daniel , Pereda Ariel
Published: November 2013

Background

Influenza A viruses (IAV) are important pathogens responsible for economic losses in the swine industry and represent a threat to public health. In Argentina, clinical, pathological, and virological findings suggest that IAV infection is widespread among pig farms. In addition, several subtypes of IAV, such as pH1N1, H3N2, δ1H1N1, and δ2H1N2, have been reported.

Objectives

To evaluate the infection patterns of influenza virus in nine pig farms in Argentina.

Methods

Clinical, serological, pathological, and virological cross‐sectional studies were conducted.

Results

Clinical and pathological results were characteristic of endemic influenza infection in eight of the nine farms studied. By rRT‐PCR, six of the nine farms were positive to influenza. Five IAV were obtained. Genome analysis determined that four of the isolations were pH1N1 and that the remaining one was a reassortant human origin H3N2 virus containing pandemic internal genes. Serological results showed that all farms were positive to influenza A antibodies. Moreover, the hemagglutination inhibition test showed that infection with viruses containing HA′s from different subtypes (pH1, δ1H1, δ2H1, and H3) is present among the farms studied and that coinfections with two or more subtypes were present in 80.5% of positive pigs.

Conclusions

Because vaccines against IAV are not licensed in Argentina, these results reflect the situation of IAV infection in non‐vaccinated herds. This study provides more information about the circulation and characteristics of IAV in a poorly surveyed region. This study provides more data that will be used to evaluate the tools necessary to control this disease.
Immunofluorescence of Japanese Eneephalitis Virus InfectionIn Vitro: Localization of Viral Antigen in Canine Cerebellar Tissue Cultures
Authors: Yoshida Jun , Hotta Susumu
Published: November 2013

Propagation of Japanese encephalitis (JE) virus in cells of dog cerebellar tissue cultures was investigated by means of fluorescent antibody (FA) technique. The fluorescent globulin conjugate was made from the serum of a dog inoculated with partially purified JE virus, treated by Sephadex G‐25 and DEAE cellulose column chromatography and then adsorbed with dog liver powder. This preparation was found to be appropriate for the present work. Fluorescence was demonstrable in virus‐infected cultures of three different types of cells, fibroblast‐like cells, nerve cells and some of the glial type cells. Fluorescence could first be demonstrated about 20 hours after virus inoculation and appeared to increase in intensity in proportion to the increase of infective virus present in the cultures. The specificity of the reaction was supported by the non‐reactivity of control (non‐infected) cultures and by the results of blocking tests. The infected nerve cells and glial type cells also exhibited morphological changes clearly detectable by the FA techniques, corresponding to the changes shown in Bodian‐stained preparations. The localization of FA antigen in the fibers of these cells suggests a possible mode of spread of JE virus in the nervous tissues. In any of the cell types studied thus far, the nuclei remained FA‐unstained even during the advanced stage of infection.
Immunobiologic Role of the Bursa of Fabricius and Thymus in the Development of the Competence to Produce Serum Antibody in Chickens
Authors: Takahashi Katsuji
Published: November 2013

Groups of chickens which were surgically thymectomized or bursectomized immediately after hatching, or hormonally bursectomized by testosterone injection at an early embryonic stage, or thymo‐bursectomized were used in an attempt to elucidate the immunobiologic roles of the bursa of Fabricius and the thymus of the chicken in the initial development of immunologic competence, particularly the ability to produce serum antibody. There were no significant differences in the rate of elimination of an injected antigen (131I‐human serum albumin) from the blood of all the groups. However, a significant difference was observed in antibody production; no antibody was produced in the sera of hormonally bursectomized birds after the primary and secondary injections, while there was recognizable production of antibody in the sera of surgically thymectomized or bursectomized birds. Further, no antibody production was observed in tissue cultures of spleen cells derived from hormonally bursectomized chickens after immunization with human serum albumin or sheep erythrocytes. The injection of viable cells of the bursa restored to a significant degree the impaired capacity to produce antibody in hormonally bursectomized and surgically thymectomized birds as well as surgically thymo‐bursectomized birds, while cell‐free extracts of the bursa could restore the capacity only in the latter group of birds. These results suggest that in the chicken, only the bursa is concerned in the development of immunologic competence as measured by the ability to produce serum antibodies. The main role of the bursa seems to be to provide the initial supply of lymphoid cells and, in addition, the production of a humoral factor which promotes the proliferation of lymphoid cells and their immunological maturity.
Relationship between Indole Production, Nitrite Production and Carbohydrate Fermentation in Vibrio cholerae
Authors: Ozawa Atsushi
Published: November 2013

Vibrio cholerae produces indole in the presence of nitrite even though the former cannot always be demonstrated as such by standard test procedures. The results of present experiments indicate that the inhibition of a positive indole reaction by nitrite may be due to the interaction of nitrite with the para‐dimethyl‐amino‐benzaldehyde in the indole reagent. Several factors, such as strain differences, pH change of culture medium, nature of fermentable carbohydrates, are also associated with the inhibition of indole production in the presence of fermentable substrates. The way in which these factors, singly or in combination, affect this inhibition is not known.
Prevalence of Transferable Drug Resistance in Drug‐Resistant Bacteria Isolated from Urinary Tract Infections in Japan
Authors: Naide Yorio , Kawamura Takeshi , Makino Kozo , Tamura Hajime , Watanare Tsutomu
Published: November 2013

Many urinary isolates which belong to the Enterobacteriaceae and bear multiple drug resistance were shown to harbor transmissible resistance factors (R factors). The levels of resistance were almost uniform in every strain investigated. The resistance markers were usually transferred as a unit. Frequency of transfer varied from host to host even when the same recipient strain was employed. However, no remarkable differences were observed in successive transfers of R factors between substrains of Escherichia coli K‐12. The role of R factors in urinary tract infection is discussed.
The Electrophoretical Analysis of Esterase and Catalase and Its Use in Taxonomical Studies of Mycobacteria
Authors: Nakayama Yasuko
Published: November 2013

The zymogram patterns of esterases and catalases of mycobacterial strains were studied using the thin layer agar electrophoresis. Though there were some variations, Mycobacterium hominis, M. bovis, M. kansasii, M. fortuitum, M. runyonii, M. avium, M. phlei and M. smegmatis seemed to show species‐specific patterns consisting of 2 to 6 esterase bands and one or more catalase bands. The patterns of scotochromogens and nonchromogens were rather variable.
Complement Fixation Reaction and Agar Gel Double Diffusion Test in Tyzzer's Disease of Mice
Authors: Fujiwara Kôsaku
Published: November 2013

Complement fixation reaction and agar gel double diffusion test were made using as antigens, water or phosphate buffer saline extracts of liver from mice severely infected with Tyzzer's disease. The supernatant of the extract heated at 56 C for 30 minutes was highly reactive with immune mouse serum in the complement fixation test, while an intense precipitate band was observed only with unhealed extracts in agar gel double diffusion tests. Cross reactions were observed in both tests between liver extracts from mice infected with different strains of organisms originating from different sources and showing varied degrees of virulence. Such reactions were not shown between Tyzzerimmune serum and liver extracts from uninfected or murine hepatitis‐infected mice, nor were they shown between the Tyzzer‐antigen and normal or anti‐murine hepatitis virus serum. The complement fixation test was used to test retired breeder mouse sera for disease contamination in mouse colonies. Positive reactions were observed in test sera collected from some breeding colonies that subsequently proved to be contaminated by the cortisone provocation test in young animals.
Grouping of RNA Phages by a Millipore Filtration Method
Authors: Miyake Tadashi , Shiba Tadayoshi , Watanabe Itaru
Published: November 2013

Twenty‐five strains of RNA phages were tested for their filtration and elution patterns (F‐E patterns) by a millipore filtration method. These strains, including MS2, f2 and R17, were separated in three groups in this aspect. We named these groups as group I, II and III. According to this grouping, MS2, f2 and R17 belonged to group I and Qβ belonged to group III. Furthermore, it was shown that group III was further divided in two sub‐groups (IIIa and IIIb) by this method. Grouping based on the F‐E patterns was in extremely good accordance with the grouping based on the serological properties. This grouping was also supported by the results of chemical and physical analyses of these RNA phages, that is, RNA phages which belonged to the same group had several common properties. Basic Information on the millipore filtration method was also presented in this paper.
A Statistical Approach to the Definition of Bacterial Species
Authors: Tsukamura Michio
Published: November 2013

A statistical method is proposed for recognition of a bacterial species or for differentiation of two groups of bacterial strains. Comparison between two groups is done by the “t”‐test. When the mean S‐values (similarity value) of two groups are A and B, and the mean S‐value for all possible combinations between strains of both groups is S, a condition necessary for defining the two groups as different species is to demonstrate the existence of equations A > S and B > S. Unless this condition is fulfilled, the two groups should be considered unseparable. The condition necessary for recognition of two groups as one species is to demonstrate the existence of equations A:= S and B = S. A few examples of the test were shown using the mycobacteria, and it was suggested that this statistical method is useful in the recognition of a species.
A Phase Cinematography Study on Morphological Changes of FL‐Line Cells Following the Infection of Poliovirus
Authors: Makino Sajiro , Nakanishi Yuh H. , Yoshida Michihiro C. , Sakurada Norio
Published: November 2013

Successive cellular changes leading to damage induced in human epithelial FL‐line cells by poliovirus infection were followed by means of phase cinematography and described on the basis of cine records of living cells and fixed‐stained specimens. The first visible change appeared as acidophilic inclusions in nuclei of stained specimens as early as 5 hours after infection. Following distortion of the nucleus, a marked increase in basophilia and the appearance of a paranuclear mass were noted in the cytoplasm. With the development of this mass, the nucleus located in the peripheral area of the cell became intensively wrinkled and distorted. Then several vacuoles appeared in the cytoplasm, and a marked blebbing of the cell surface followed, resulting in shrinkage of the cell. The cell damage was characterized morphologically 20 to 24 hours after infection by an eosinophilic cytoplasmic mass, severe distortion of the nucleus and fragmentation of nuclear material leading to pycnotic degeneration of nuclei. The nucleoli and acidophilic intranuclear inclusions remained unchanged during the course of pyknotic degeneration of cells. Degenerative changes leading to cell damage proceeded rather slowly in infected cells.
Fine Structure of Strictly Anaerobic, Non‐Spore‐Forming, Gram‐Negative Bacilli
Authors: Ushijima Tsutomu
Published: November 2013

The fine structure of a strain of Bacteroides insolitus has been studied by ultrathin sectioning and electron microscopy. Logarithmically growing cells were fixed both by osmic acid and potassium permanganate, and embedded in Epon. Thin sections were stained with uranyl acetate and examined. The periphery of the cell was composed of a wavy three‐layered outer membrane (ca. 80 A), an intermediate layer (50–200 A), and three layered cytoplasmic membrane (ca. 80 A). Single or double bridges which connected the outer membrane with the cytoplasmic membrane were observed. Invagination of the cytoplasmic membrane was observed in no occasion. Independent, distinct, and uniform particles were not the main component of the cytoplasm. The cytoplasm was filled with more or less beaded reticulum‐like structures. The nucleoplasm with fine fibrils was mainly dispersed continuously in rather regular cubic masses in an intermediate region between the center and the periphery of the cell. Contacts of the nucleoplasm with the cytoplasmic membrane were occasionally observed.
Studies on the Protection Test of Pertussis Vaccine. Immunization Period and Protectivity
Authors: Yoshioka Morimasa , Takatsu Kuniyoshi , Kawahira Minoru , Takahashi Kihachiro
Published: November 2013

In order to confirm the data reported in the previous papers, variously prepared pertussis vaccines were employed in the present investigation. Pertussis organisms grown either on a solid or in a liquid semisynthetic medium were treated by: (1) heating at 56 C for 30 min, (2) storage in 0.1% formalin at 37 C for 5 days, (3) storage in 0.1% formalin at 25 C for 5 days, and (4) simple addition of sodium ethyl‐mercuri thiosalicylate (merthiolate) as a preservative. Freeze‐dried vaccines were made from these preparations four and a half months later. Mice were immunized intraperitoneally and challenged intracerebrally with a virulent strain of Bordetella pertussis 10 or 17 days later. The data were analyzed statistically assuming the probit corresponding to the percentage of survivors at any dose be a linear function of the logarithms of the dose. The 50% effective doses (ED50) and slopes of each vaccine were found to be uniform. More accurate estimates of ED50 were obtained by employing a pooled slope in each experiment. From these ED50 values, the relative potency was estimated by comparing the value of a vaccine to that of a dried merthiolate‐vaccine. For vaccines derived from solid cultures, with an immunization period of 17 days, the relative potency of the vaccine heated at 56 C was 0.63 (95% fiducial limits=0.52 to 0.76); the value for the formalinized vaccine at 37 C was 0.40 (0.30 to 0.53) and one at 25 C was 0.51 (0.34 to 0.77). Vaccines derived from liquid cultures showed a relative potency of 20 to 50% less than that of corresponding vaccine derived from a solid culture. The potency obtained for the 17 day immunization period was usually higher than that for the 10 day period. Using the overall‐pooled slope, an experimental design which will be appropriate for statistical analysis is discussed.
Penicillin Resistance Correlated with Phage Types in Staphylococcus aureus
Authors: Fujisawa Hiroyasu , Ishihara Keizo
Published: November 2013

The resistance of Staphylococcus aureus to penicillin G was studied quantitatively by the dilution method. A series of 338 strains obtained from patients fell into 3 groups according to degree of resistance; 150 highly resistant strains, 137 moderately resistant strains, and 51 sensitive strains. Furthermore, 287 strains resistant to 0.2 units per ml or more of penicillin fell into 2 groups according to antibiotic resistance pattern, 173 single resistant strains and 114 multiple resistant strains. The vast majority of the highly resistant strains belonged to phage‐groups I and III, whereas the majority of the moderately resistant strains belonged to group II. Of the highly resistant strains, 65% were multiple resistant, and 35% single resistant. Of the multiple resistant strains, 86% belonged to the 52, 52A, 80, 81 complex within group I and to group III, but none belonged to group II. Of the strains that were highly resistant to penicillin only, 83% belonged to group I, and of these, nearly one‐half were members of the 52, 52A, 80, 81 complex, and the other half belonged to the rest of phage‐types within group I. In contrast, 88% of the moderately resistant strains were single resistant, and of these, 58% belonged to group II. Mercury resistance was observed in 77% of the highly resistant strains of the multiple resistance pattern, 29% of those of the resistance to penicillin alone and 3% of the strains moderately resistant to penicillin alone. Of the mercury resistant strains, 68% belonged to the 52, 52A, 80, 81 complex, 3% to the rest of phage‐types in group I, 12% to group III, but none belonged to group II. The results suggest that there may exist a difference in the mechanisms of inactivating penicillin between strains of staphylococci highly resistant and moderately resistant to penicillin.
Serological and Pyocine Types of Pseudomonas aeruginosa from Various Sources
Authors: Matsumoto Hideki , Tazaki Tadakatu , Kato Takashi
Published: November 2013

Two hundred and sixty six strains of Pseudomonas aeruginosa isolated from various pathological or non‐pathological materials were classified by serological and pyocine typing methods for the purpose of epidemiological investigation. The O antigenic groups detected were Verder and Evans' O1, 2, 3 (7a, 7b), 4, 5, 6, 8, 9, 10 and Habs' 04 and O10. In addition, three new O antigenic groups designated tentatively as Ox11, Ox12 and Ox13 have been established. Among these groups, Verder and Evans' O2, O3 (7a, 7b) and Ox11 were most common. No significant difference existed between the O groups of P. aeruginosa isolated from pathological materials and those obtained from non‐pathological sources. Pyocine typing of Darrell and Wahba was considered to be useful in the epidemiological investigation. In a patient, P. aeruginosa of a specific sero‐pyocine type persisted for long period in an infected focus.
Complement‐Fixation Tests of Rickettsia orientalis with Sodium Deoxycholate Treated Antigen
Authors: Tachibana Nobuyoshi , Kobayashi Yuzuru
Published: November 2013

Complement‐fixation tests of Rickettsia orientalis were performed by using antigens purified with sodium deoxycholate. Cross complement‐fixation tests between antigens of the Gilliam, Ohzeki, Karp and Kato strains and immune mouse sera of these four strains and of the Kaneko strain revealed difference in antigenic structure among the Gilliam, Karp and Kato strains. The Ohzeki strain was identical with the Gilliam strain and the Kaneko strain was identical with the Karp strain. In the homologous system, complement‐fixing antibodies of mice infected with R. orientalis began to appear on the 15th day after infection and rose rapidly in a few days, and reached maximum in 4–6 weeks after infection and 8 weeks at latest. Subsequently, the titers declined gradually and showed low titers on the 52nd week in spite of survival of rickettsia in mice. In the heterologous system among which antigenic structures were different, the titers rose somewhat later and declined earlier than those of the homologous system. It was confirmed that the antigen treated with sodium deoxycholate was utilizable in the specific serological diagnosis of scrub typhus, because convalescent sera from all ten patients with the disease showed significant rise of complement‐fixing titers against the antigen.
Biological Activity of Papain Digested Streptococcal Anti‐M Antibody
Authors: Saito YÜIchi
Published: November 2013

Digestion of rabbit streptococcal anti‐M antibody with papain to produce univalent fragments resulted in the loss of its ability to fix complement. However, the digested univalent antibody still retained the indirect bactericidal activity in vitro, opsonic activity in vivo, and also the capability of protecting mice against the streptococcal infection. Thusly the biological activities of digested anti‐M antibody appear to be similar to those of intact anti‐M antibody.
Heat Resistance and α‐Toxigenicity of Clostridium perfringens Strains in Normal Intestines of Japanese
Authors: Nakagawa Masaaki , Nishida Shoki
Published: November 2013

Heat‐resistant strains of Clostridium perfringens were isolated from normal feces samples of Japanese people more frequently than from European people. However, the predominant number of the C. perfringens cells in the human intestines were sensitive to the heat at 100 C for 10 min. This proved to be true of the feces samples which were demonstrated to carry the organism resistant to 100 C for 60 min. Further studies indicated that the more the number of the organism in the intestines, the higher was the recovery ratio of the heat‐resistant strains. Concomitant estimation for α‐toxigenicity of those strains isolated from unheated feces samples revealed that the α‐toxigenicity ranged mainly between 0.05 and 0.9 α‐antitoxin equivalents.
Ibaraki Virus, an Agent of Epizootic Disease of Cattle Resembling Bluetongue
Authors: Omori Tuneyoshi , Inaba Yuji , Morimoto Tomiaki , Tanaka Yoshio , Ishitani Ruizo , Kurogi Hiroshi , Munakata Kozo , Matsuda Kazuo , Matumoto Minoru
Published: November 2013

Outbreaks of an acute febrile disease of cattle occurred in Japan in 1959 and 1960. Its occurrence was limited in late summer and autumn, and in Kyushu, Shikoku and Honshu roughly south of 37 degrees north latitude, suggesting a close correlation of the incidence with the climatic conditions, hence a possibility of the presence of arthropod vector. The disease was characterized by fever and lesions affecting the mucous membrane and skin, musculature and vascular system. Degeneration of striated muscles was observed in the esophagus, larynx, pharynx, tongue and skeletal muscular system. Edema and hemorrhage were marked in the mouth, lips, abomasum, coronets etc., occasionally followed by degeneration of the epithelium leaving erosions or ulcerations. Severe lesions affecting the esophageal and laryngopharyngeal musculature caused deglutitive difficulty which in turn resulted in dehydration and emaciation, and occasionally in aspiration pneumonia, constituting the major causes of death of the affected animals. These findings indicate that the disease resembles bluetongue in sheep and cattle. The clinical materials obtained from natural cases induced a clinical illness when inoculated into calves, and the disease was transmitted serially in calves by intravenous inoculation of the blood obtained at the height of febrile reaction. The experimentally produced disease was clinically and pathologically indistinguishable from the natural disease.
Drug Resistance of Staphylococci
Authors: Mitsuhashi Susumu , Kono Megumi , Sagawa Mozo , Mori Hiromichi
Published: November 2013

Induction of chloramphenicol (CM) resistance in Staphylococcus aureus was investigated by using several CM derivatives. It was found that dl‐threo‐1‐p‐nitrophenyl‐2‐dichloroacetamino‐1,3‐dichloropropane has high antibacterial activity but low activity of induction for CM resistance. In spite of low antibacterial activity, induction of CM resistance occurred after prior treatment with dl‐threo‐1‐p‐nitrophenyl‐2‐dichloroacetamino‐3‐chloropropane‐1‐ol. It was found that dl‐chloramphenicol di‐acetate, dl‐threo‐1‐p‐nitrophenyl‐2‐dichloroacetamino‐3‐bromopropane‐1‐ol and dl‐threo‐1‐phenyl‐2‐dichloroacetamino‐1,3‐propanediol have induction ability in spite of the absence of antibacterial activity. Other derivatives were classified into two groups; (1) low antibacterial activity and induction of CM resistance and (2) loss of both activities.
Distribution of R Factors among Shigella Strains Isolated in Japan
Authors: Tanaka Tokumitsu , Nagai Yutaka , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

Shigella strains isolated in Japan from 1965 to 1967 were surveyed for drug resistance and distribution of R factors. Of 5,875 strains tested, 5,429 were resistant to either one or various combinations of four drugs, tetracycline (TC), chloramphenicol (CM), streptomycin (SM) and sulfanilamide (SA). Among these resistant strains, 80.3, 1.8, 2.2, and 15.7% were quadruply, triply, doubly and singly resistant, respectively. Fifty percent of these resistant strains were found to carry R factors which are capable of conjugal transfer. More than 50% of TC or CM resistant strains carried R factors, but in the remaining resistance type, R factors were rarely isolated. Only 3 and 17% carried R factors among SA and SM.SA resistant strains, respectively. The strains resistant to drugs other than the aforementioned four were very few; 0.9, 0.7 and 0.01% being resistant to aminobenzyl‐penicillin (APC), nalidixic acid (NA) and kanamycin (KM), respectively. Conjugally transmissible NA resistance was not found among strains so far isolated and examined. About one third of APC resistance was transmissible. Among 9,635 strains examined, one R factor possessing the KM resistance marker and 28 R factors possessing the APC resistance marker were demonstrated. The R factor possessing the KM resistance marker was multiply resistant to 8 drugs; TC, CM, SM, SA, APC, KM, fradiomycin and paromomycin.
Integration of Chloramphenicol Resistance Gene of an R Factor on Escherichia coli Chromosome
Authors: Iyobe Shizuko , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

An unstable mutant R factor conferring only chloramphenicol (CM) resistance was obtained by spontaneous segregation. After storage in broth culture, a stable CM‐resistant mutant was obtained and its CM‐resistance could not be cured by treatment with acriflavine or transduced to a recombination‐deficient strain of Escherichia coli K12. Recombinational analysis indicated that the cml gene governing CM resistance had been integrated into the E. coli chromosome and closely linked with met B locus. The cml gene was co‐transduced with both met and arg markers by phage P1, and the linkage order was considered to be mtl‐cml‐met‐arg‐thi. When the strain carrying this chromosomal CM‐resistance was infected with a transferable R (TC) factor capable of conferring tetracycline (TC) resistance, the CM‐resistance became transferable by conjugation. This mechanism is considered to account for the formation of the recombinant R (TC.CM) factor.
Demonstration of R Factors in Enteric Bacteria Isolated at the Boston City Hospital
Authors: Egawa Ryuki , Hara Yoshihiko , Mitsuhashi Susumu , Cohen Stephen N.
Published: November 2013

Fifty‐one strains of enteric bacteria isolated in 1966 from patients at the Boston City Hospital were studied for the distribution of R factors. Twenty strains were found to carry transmissible drug‐resistance factor “R”. The strains harboring R factors included Escherichia coli, Aerobacter, Klebsiella, Proteus and Serratio. This report confirms similar studies in suggesting that R factors are widely distributed in enteric bacteria isolated in the United States.
Immunoglobulin Classes of Serum Antibodies Formed in Response to Natural Infections with Myxo‐ and Pseudomyxoviruses among Infants and Children
Authors: Sekiguchi Hiroshi , Numazaki Yoshio , Yano Namio , Homma Morio , Ishida Nakao
Published: November 2013

The occurrence of immunoglobulin classes of serum antibody in response to natural infections with myxo‐ and pseudomyxoviruses among infants and children was investigated. Two types of the 19S antibody response have been demonstrated. In the first type, in which are included the measles and mumps virus infections, the 19S antibody appeared dominantly as a primary response to the virus infection. In the second type, which includes the influenza, parainfluenza and RS virus infections, the 19S antibody was either not detectable or produced in a limiting amount in infants even at an early stage of the illnesses. A possibility was discussed that production of the 19S antibody in response to the infection with myxo‐ and pseudomyxoviruses may be primarily determined by the nature of virus infection itself, particularly the presence or absence of a viremia.
Experimental Salmonellosis Immunizing Effect of Live Vaccine Prepared from Various Mutants of Salmonella Having Different Cell Wall Polysaccharides
Authors: Kawakami Masaya , Nakata Hisako , Mitsuhashi Susumu
Published: November 2013

Immunizing potencies of vaccines prepared from various strains of Salmonella were graded by comparing the mortality rate of immunized mice after challenge with highly virulent strains of either Salmonella enteritidis or S. typhimurium. The resistance against this challenge infection was shown to be conferred by joint immunization with a specific factor, which was represented by O specific lipopolysaccharide of smooth strains, and cross‐protection factor, which was a major potent factor in live vaccine. The distribution of this cross‐protection factor in rough mutants of S. typhimurium was found to be limited to strains which possessed a polysaccharide chain longer than that of glucose1‐less mutant. The potency conferring cross‐resistance was found to be maintained partly in formalin‐killed cells and cell walls of the strains harboring cross‐protection factor but not in lipopolysaccharide extracted from such strains.
Studies on the Antigenic Activities of Yeasts
Authors: Sunayama Hiroyoshi
Published: November 2013

In order to identify the antigenic determinant groups of the mannan of C. albicans serotype A, six kinds of manno‐oligosaccharides of up to 7 units in chain‐length connected by α1→2 linkages were prepared from the partial acetolysate of the parent mannan. In the precipitation‐inhibition test of anti‐C. albicans serotype A serum with its homologous mannan, inhibitory power of the oligosaccharides was of the following order: heptaose→: hexaose>pentaose>tetraose>triose> biose, and the amounts for 50%‐inhibition of the former four oligosaccharides were 0.08, 0.10, 0.50 and 3.0 μmole respectively, and the inhibitory power of the latter two oligomers at 0.5 μmole were 8 and 5% respectively. On the other hand, the cross‐inhibition test of anti‐C. albicans serotype A serum with the heterologous mannan of C. albicans serotype B afforded the result that the order of inhibitory activities was hexaose>heptaose>pentaose>tetraose>triose> biose, and that the amounts for 50%‐inhibition were 0.05, 0.08, 0.1, 0.45, 0.50 and 3.0μmole respectively. Furthermore, the results of inhibition test on the anti‐C. albicans serotype A serum absorbed with the mannan of C. albicans serotype B revealed that the biose, triose and tetraose did not show significant inhibitory power in the range employed, whereas the pentaose, hexaose and heptaose did not significantly affect the inhibitory activities. Thus, it was concluded that the antigenic determinants of the mannan of C. albicans serotype A are α1→2 linked hexaose or heptaose moieties. Based on the above facts, the serological differences between two antigenic mannans of C. albicans serotype A and B may reside at least in the length of the antigenic determinants in which the former is longer than the latter considering the length of the α‐D‐manno‐pyranosyl residue.
Transfer Agent of Immunity
Authors: Kurashige Satonori , Kitamura Kouichi , Akama Kiyoto , Mitsuhashi Susumu
Published: November 2013

An immune ribonucleic acid (RNA) preparation was extracted with phenol from the spleens of guinea pigs immunized with diphtheria toxoid. Antibody‐carrying cells were detected by immunocyte adhesion as rosette‐forming cells. When germ‐free rats, conventional guinea pigs or mice were injected intraperitoneally with this preparation, the rosette‐formers were detected in either peritoneal exudate cells or spleen cells, whereas serum antibodies were unable to be detected thus far in such animals. Two injections with this preparation did not cause any remarkable increase in the number of rosette‐formers, and serum antibody was also not detectable. By contrast, a high titer of serum antibody was demonstrated and the number of rosette‐formers increased shortly after an injection of a small amount of diphtheria toxoid into guinea pigs which had previously received an injection with immune RNA. This reaction indicates a secondary response of antibody formation. However, secondary responses were not induced by injections of immune RNA preparations in guinea pigs primed with either diphtheria toxoid or immune RNA preparation. These facts suggest that immune RNA preparations did not contain antigens or fragments thereof and the immune response induced by RNA preparation is not the same as that induced by stimulation by the antigen itself. These results moreover can be accounted for by the notion that the immune RNA preparation is able to induce “memory” cells capable of responding to a secondary stimulus with an antigen and producing a high titer of serum antibody.
Nalidixic Acid Cetrimide Agar
Authors: Goto Sachiko , Enomoto Seiji
Published: November 2013

Attempts were made to modify the formula devised by Brown and Lowbury (1965) for more effective selection of Pseudomonas aeruginosa. An improved formula was devised by the authors as follows: Peptone (Eiken) 2.0%, dipotassium phosphate 0.03%, magnesium sulfate (anhydrous) 0.03%, cetrimide 0.02%, nalidixic acid 15 μg/ml, agar 1.5%, with a final pH 7.4–7.6. On this media the growth and pigment production of Ps. aeruginosa was nearly equal to that of Brown‐Lowbury's composition. Strains of Enterobacteriaceae such as Klebsiella, Proteus and Providencia were strongly suppressed on the author's medium although some of them showed a moderate growth on Brown‐Lowbury's medium.
Determination of Pseudomonas aeruginosa by Biochemical Test Methods
Authors: Arai Taketoshi , Enomoto Seiji , Goto Sachiko
Published: November 2013

A new synthetic medium consisting of 0.2% KH2PO4, 0.01% MgSO4·7H2O, 0.5% NaCl, 0.05% (NH4)2SO4 and 1.0% Tween 80 was devised by the authors as a means of differentiating between Pseudomonas aeruginosa and related species of gram‐negative rods. One loopful of an overnight nutrient agar culture of a test strain was inoculated into 1 ml of this media, and incubated at 37 C for 24 hr. All of the 38 test strains of P. aeruginosa showed visible growth in this media, being capable of utilizing Tween 80 as a sole carbon source, whereas strains of Aeromonas, Vibrio parahaemolyticus, Achromobacter sp., Proteus sp., Escherichia coli and Citrobacter sp. could not grow within a 24 hr incubation period, although longer incubation yielded a visible growth with some strains. A strain of P. fluorescens did not show any visible growth at 37 C, but did show a moderate growth after a 48 hr incubation at 18 C. Strains of Klebsiella and Enterobacter showed only slight growth after incubation at 37 C for 24 hr.
RNA‐Induced Interference with Animal Virus Infection
Authors: Matsuyama Masako , Kawade Yoshimi , Fukada Tetsuo
Published: November 2013

Some RNAs, including both single‐ and double‐stranded RNAs, when incubated with chick embryo cell culture induce cellular resistance against viruses. Evidence was now obtained indicating that the induction of cellular resistance by RNA depends on the cellular metabolic activity, especially on the synthesis of cellular RNA and protein. Thus, inhibitors of RNA and protein synthesis, actinomycin D and cycloheximide, were found to inhibit the development of an antiviral state when added before, or during the relatively early period of, incubation of the cells with RNA. In the course of induction of cellular resistance, three stages may be distinguished, the priming stage, the developing stage, and the established resistant stage.
Transfer Agent of Immunity
Authors: Saito Kazuko , Osawa Nobutaka , Mitsuhashi Susumu
Published: November 2013

An immune ribonucleic acid (RNA) was extractable from the spleen cells of mice hyperimmunized with live vaccine of Salmonella enteritidis. The RNA was capable of inducing cellular immunity and developing cellular antibody in the peritoneal macrophages of mice injected with this agent. It was found that cellular immunity was detectable even 90 days after injection in the peritoneal macrophages of mice which had received an intraperitoneal injection with this agent. Results of serial passive transfers of cellular immunity through immune RNA led us to the conclusion that this agent does not contain antigen or fragment thereof and may replicate actively in the recipient cells, although the mechanism still remains to be elucidated. The development of cellular immunity by immune RNA was inhibited by puromycin but not by actinomycin D. However, serial passive transfers of cellular immunity through immune RNA was inhibited by treatment of recipient mouse with actinomycin D, implying the role of DNA‐dependent RNA polymerase in the processing of immune RNA in recipient cells. Using these results, the role of immune RNA and the possible mechanisms of immune RNA replication are discussed.
Purified K‐Agglutinogen of Bordetella pertussis and Its Properties
Authors: Nakase Yasukiyo , Kasuga Tadayoshi
Published: November 2013

We attempted to purify the K‐agglutinogen of Bordetella pertussis by mild procedures. From the supernatant of sonic treated cells, the K‐agglutinogen was separated by a successive use of O‐diethylaminoethyl (DEAE)‐cellulose column chromatography, ammonium sulfate precipitation and O‐carboxymethyl (CM)‐cellulose column chromatography. The specific activity of purified K‐agglutinogen increased 64–128 fold without damaging either the L or S antigens, and recovery was about 86%. A fraction containing mainly S‐agglutinogen, was also obtained. The K‐agglutinogen, which was a simple protein with an S value of 2.5, was confirmed to be in a highly purified state by a gel‐diffusion test, K‐antibody production and physicochemical tests, and was also found to be free from other biologically active substances of B. pertussis: namely heat labile toxin, hemagglutinin, histamine‐sensitizing factor and O‐antigen. No protective activity was found in the purified K‐agglutinogen or against intracerebral challenge with B, pertussis in mice.
Mycobacterium rhodesiae sp. nov.
Authors: Tsukamura Michio , Mizuno Shoji , Gane N. F. F. , Mills A. , King L.
Published: November 2013

A number of rapid‐growing scotochromogenic mycobacteria were isolated from the sputum specimen of Rhodesian patients with pulmonary disease and recognized as a new species. This was given the name Mycobacterium rhodesiae sp. nov. A comparison between M. rhodesiae, M. parafortuitum, M. aurum and M. vaccae was done, and distinguishing characters serving for differentiation between these 4 species of rapid‐growing scotochromogenic mycobacteria were described.
The Germination Requirements of Spores of Clostridium botulinum Type E
Authors: Ando Yoshiaki
Published: November 2013

The requirements for, and conditions influencing, the germination of spores of Clostridium botulinum type E were studied in chemically defined media. Spores of this organism germinated rapidly and completely in various combinations of L‐alanine and one of such carbon sources as sugars, lactate, and ribosides in the presence of bicarbonate. The most effective compounds tested were glucose, L‐ and D‐lactate, and inosine. The formation of lactic acid was found to occur during germination of spores in a combination of L‐alanine and glucose. Furthermore, the germination in this medium was strongly inhibited by fluoride or iodoacetate. The germination of spores was also initiated with high concentrations (100 mM or more) of L‐alanine or certain other amino acids alone in the presence of bicarbonate at pH 9.0. The spores germinating in L‐alanine alone and those in L‐alanine plus other compounds were compared as to the minimal concentration of L‐alanine required, pH optimum, temperature optimum for heat‐activation, bicarbonate requirement, and effects of various inhibitors. Significant differences among these characteristics led to a conclusion that different metabolic pathways, operating selectively each germinative compound used, might be involved in the germination of spores of the organism. The mode of action of combination in stimulating germination by the different germinative compounds has been discussed.
The Multiplication of Mycobacterium leprae in Mouse Foot‐Pads and the Ambient Temperatures
Authors: Matsuo Yoshiyasu
Published: November 2013

The influence of fluctuation of the environmental temperatures on the multiplication of Mycobacterium leprae in mouse foot‐pads was investigated. After infection with 104 M. leprae into the foot‐pads, mice were divided into the following 4 groups: Group 1, mice were housed in a room maintaining the air temperature at 20 C with thermoregulatory devices throughout the experiment; Group 2, kept at 20 C for the first 16 weeks, and at a regular animal room where the air temperatures fluctuated seasonally and diurnally for the next 15 weeks and then returned to the 20 C room; Group 3, kept at 20 C for the first 16 weeks and thereafter at the regular animal quarters; Group 4, kept at the regular animal room throughout the experiment. In the mice in Group 1, M. leprae increased as expected. Acid‐fast bacilli also multiplied in the mice kept under fluctuation of air temperatures at the regular animal quarters but distinctly more slowly than in those kept at 20 C. Acid‐fast bacilli rose in the mice transferred to the 20 C room from the regular ones, suggesting that viability of M. leprae is not significantly affected by ordinary fluctuation of air temperatures.
Specific Establishment of Lactobacilli in the Digestive Tract of Germ‐Free Chickens
Authors: Morishita Yoshiyuki , Mitsuoka Tomotari , Kaneuchi Choji , Yamamoto Shutaro , Ogata Manabu
Published: November 2013

Establishment of lactobacilli in the digestive tract of germ‐free chicks was studied using the organisms isolated from chickens, human infants and sour milk and those received from the American Type Culture Collection. In the case of monocontamination, intestinal lactobacilli such as Lactobacillus acidophilus, L. salivarius and L. fermenti except L. acidophilus ATCC 4356 and non‐intestinal lactobacilli such as L. plantarum and L. casei were well established in the digestive tract of germ‐free chicks regardless of their origins, although the bacterial numbers of the established strains varied considerably in different parts of the gut. On the contrary, L. acidophilus ATCC 4356 of human origin and L. jugurti, L. helveticus and L. brevis which originated from non‐intestinal materials failed to be established in the gut of germ‐free chicks. In the case of dicontamination, L. acidophilus from an infant was shown to disappear after administration together with Escherichia coli or L. salivarius from chickens. L. plantarum and L. casei disappeared or showed a suppressed growth in the presence of L. acidophilus or Streptococcus faecalis var. liquefaciens from chickens.
The Augmenting Effect of Antibody on the Induction of an Immune Response
Authors: Hamaoka Toshiyuki , Takatsu Kiyoshi , Kitagawa Masayasu
Published: November 2013

Primary and secondary antigenic stimuli were given in the presence of antibody in order to study the role of antibody in the induction of the subsequent immune response. Primed cells immunized with crystalline bacterial α‐amylase (BαA) and crystalline Taka‐amylase A (TAA) were more effectively stimulated by antigen bearing cells which had been previously prepared by exposure to the two antigens in the presence of anti‐BαA or anti‐TAA than in the absence of the antibodies. The augmenting effect of the antibody was specific in relation to the antibody added. The primary antibody responses to BαA and TAA were also specifically augmented by the presence of the corresponding antibody, when two antigens were mixed with the antibody and injected intravenously into normal mice. When dinitrophenyl (DNP) residues were introduced to BaA, the immune response stimulated by DNP‐BαA was also augmented by anti‐DNP antibody. Although specificity of the response augmented by anti‐hapten or anti‐carrier antibody may be determined on a cellular level, the specificity of response augmented by either of these antibodies extended to the hapten as well as the carrier‐determinants. Thus, the presence of antibody directed against any of determinants on an antigen, might result in widening the spectrum of responsiveness of the animal to other antigenic determinants.
Anti‐Vaccinia Activity of γ‐Thiochromanone‐1‐Thiosemicarbazone In Vitro and In Vivo
Authors: Tsunoda Akira , Miyazaki Keizo , Aota Toshinao , Matsumoto Shigehiro , Kumagai Katsuo , Ishida Nakao
Published: November 2013

A derivative of thiosemicarbazone, γ‐thiochromanone‐l‐thiosemicarbazone (SN‐13), which differed from N‐methylisatin‐β‐thiosemicarbazone (marboran) in that the carbonyl group in the C2 position of N‐methylisatin was lacking, has been found to possess an anti‐vaccinia effect as determined by the pulp disc method of plaque inhibition and by inhibition of cytopathic effect in tube cultures of chick embryo cells as well as by prevention of mouse tail lesions by the vaccinia virus. In tube cultures, SN‐13 was shown to be effective even when the treatment was started as late as 8 hr after virus infection, whereas no activity was observed with marboran when started from the 8th hr. SN‐13 was as effective as marboran on cross treatments of vaccinia virus with the two compounds in tube cultures, either by treatment at an early or a late stage of the virus growth. Moreover, the inhibitory effect of SN‐13 on vaccinia virus growth was completely reversed by actinomycin D similar to that observed with marboran in tube cultures. No additive effect of the two compounds was observed in animal tests.
Segregation of Unselected Markers in Mycobacterial Recombinants
Authors: Mizuguchi Yasuo
Published: November 2013

Analysis of linkage and segregation of unselected markers was made using a recombination system of Mycobacterium smegmatis strains, Jucho and Lacticola. It was demonstrated that recombinants primarily possessed the genetic characters derived from Jucho. This suggested that strains Lacticola and Jucho did not play equivalent roles in zygote formation, and that zygotes were made up essentially of the chromosome from Jucho and contained only a segment of the chromosome from Lacticola. The ordering of the genes in strain Lacticola might be as follows; φrb, gly‐2+, leu‐7+, and str‐9. Results of backcrosses of recombinants with parental strains showed that there were four different mating types in the recombinants. This indicated that at least two different factors controlled mating in mycobacteria. D4 phage and phage Rabinowitschi did not have mating type specificities.
Morphological Changes in Staphylococcus aureus and Escherichia coli Exposed to Cephalexin
Authors: Nishino Takeshi , Nakazawa Shozo
Published: November 2013

The action of cephalexin against Staphylococcus aureus No. 50774 and Escherichia coli NIH had been studied by phase contrast microscopy and electron microscopy. S. aureus grown on the cephalexin‐containing agar exhibited abnormal cell division when observed under the phase contrast microscope. Exposure of E. coli to cephalexin resulted in the formation of long filaments and spheroplasts. The degree of morphological changes was found to be dependent on the concentration of cephalexin added. Electron microscopic studies revealed that S. aureus upon exposure to cephalexin produced a swelling of the cross wall, which resulted in lysis of the cell or the production of a protoplast. E. coli exposed to cephalexin produced long filamentous forms. A number of mesosomes were seen in the filamentous forms. However, the presence of a cross wall or plasma membrane partitioning the elongated cell thus formed was never observed. The above‐mentioned findings offer morphological evidence that the antibacterial action of cephalexin is ascribed to the inhibition of cell division and possibly cell wall synthesis.