Endocarditis in chickens caused by subclinical infection of Streptococcus gallolyticus subsp. gallolyticus

Streptococcus gallolyticus subsp. gallolyticus causes endocarditis in humans and acute septicemia in domestic birds. We describe here the infective endocarditis caused by the bacterium found among clinically healthy broilers at two abattoirs in Japan. The chickens were thought to be healthy because of the lack of clinical symptoms and normal levels of mortality before slaughtering. At the time of inspection, some chickens were condemned because of organ disorders characterized by vegetative valvular endocarditis as well as focal necrosis in heart, liver, and spleen. Streptococcus gallolyticus subsp. gallolyticus was isolated from the organs as a pure culture, indicating that the bacterium probably was the causative agent of the disorders. Amplified fragment length polymorphism analysis of the isolates collected at the abattoirs from chickens grown in nine different farms indicated that the isolates were different variants of the same clonal lineage and may have been derived from the same ancestor. These results suggest that S. gallolyticus subsp. gallolyticus causes infectious endocarditis in chickens and that healthy chickens may possess the bacterium in their normal flora as an opportunistic pathogen.     

Isolation and characterisation of Rhodococcus equi from submaxillary lymph nodes of wild boars (Sus scrofa)

Rhodococcus equi has been isolated from the submaxillary lymph nodes of domesticated pigs, but little is known about the presence of R. equi in wild boars. The aim of the study was the evaluation of the incidence of R. equi in wild boars and the characterisation of them. Of 482 submaxillary lymph nodes of wild boars shot in 39 settlements throughout Hungary, R. equi was isolated from 60 specimens, and plasmid types of 82 isolates were examined. The isolates were tested for the presence of 15-17-kDa (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes by polymerase chain reaction (PCR). Plasmid DNAs were isolated and analysed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. None of the 82 isolates contained vapA gene but 21 isolates (25.6%) were positive for vapB gene showing 827bp product of the expected size in the PCR amplification. Sixty-one strains (74.4%) did not contain plasmid. The 21 isolates of intermediate virulence contained virulence plasmids that were identified as types 1 (1 isolate), 5 (16 isolates), 21 (1 isolate), and three new distinct plasmid variants (1-1-1 isolate), respectively. On the basis of restriction digestion patterns of plasmid DNAs, we tentatively designated the new variants as types 25-27, respectively. The prevalence of R. equi strains of intermediate virulence among the isolates originated from the submaxillary lymph nodes of wild boars (25.6%) is very similar to those of domestic pigs (26.8%) in Hungary, and plasmid type 5 is the predominating one in both groups. This is the first report of isolation of VapB-positive R. equi from wild boars in the world.     

A survey of amoebic infections and differentiation of an Entamoeba histolytica-like variant (JSK2004) in nonhuman primates by a multiplex polymerase chain reaction

A pathogenic Entamoeba histolytica-like variant (JSK2004 strain) with genetic variations and a novel isoenzyme pattern isolated from a De Brazza's guenon in a Tokyo zoo in Japan has previously been documented. In this study, a multiplex polymerase chain reaction (PCR) assay that could distinguish the JSK2004-type E. histolytica-like variant (JSK04-Eh-V) from E. histolytica and Entamoeba dispar using three newly designed primer sets for amplifying each specific DNA fragment from their small-subunit ribosomal RNA genes was developed and established. Forty-seven primates (11 species) from the zoo were surveyed by multiplex PCR to assess the prevalence of JSK04-Eh-V infection, which was recognized in six individuals of four species, including an Abyssinian colobus monkey, a De Brazza's guenon (including the individual from whom JSK2004 was isolated), a white-faced saki, and a Geoffroy's spider monkey. In addition, the autopsied individuals of an Abyssinian colobus and Geoffroy's spider monkey that died of amoebic liver abscess were also evaluated. DNA samples were also analyzed for specific genotypes based on the nucleotide sequencing of two protein-coding (chitinase and serine-rich E. histolytica protein) genes and the protein-noncoding locus 1-2 that was used for fingerprinting of the E. histolytica strain. These studies indicated that the E. histolytica-like variant infection in this zoo was caused by the same type (i.e., JSK04-Eh-V). An axenic culture medium (yeast extract-iron-maltose-dihydroxyacetone-serum) was developed based on the yeast extract-iron-gluconic acid-dihydroxyacetone-serum medium, which is designed for axenic culture of E. dispar. This new medium could be used for axenically culturing E. histolytica, JSK04-Eh-V, and E. dispar in a single medium.     

Epizootiological survey of Trichinella spp. infection in carnivores, rodents and insectivores in Hokkaido, Japan

In order to evaluate the present epidemiological situation of Trichinella infection in wild animals in Hokkaido, Japan, red foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) , brown bears (Ursus arctos) , martens (Martes melampus), rodents and insectivores captured in Hokkaido were examined for muscle larvae by the artificial digestion method from 2000 to 2006. Foxes (44/319, 13.8%), raccoon dogs (6/77, 7.8%) and brown bears (4/126, 3.2%) were found to be infected with Trichinella larvae and all other animal species evaluated were negative. Multiplex PCR and DNA sequencing revealed that larvae from a fox captured in Otofuke, in south-eastern Hokkaido, were T. nativa, and larvae from 27 animals including 21 foxes, 2 raccoon dogs and 4 brown bears captured in western Hokkaido were Trichinella T9.     

Ovsynch plus CIDR protocol for timed embryo transfer in suckled postpartum Japanese Black beef cows

We compared synchronization and pregnancy rates, and the increase in blood progesterone concentrations during luteal development, between (1) Ovsynch plus an intravaginal controlled internal drug release (CIDR) device protocol followed by timed embryo transfer (timed ET), and (2) a conventional estrus synchronization method using PGF(2 alpha) and ET in suckled postpartum Japanese Black beef cows. Cows in the PGF group (n=18) received a PGF(2 alpha) analogue when a CL was first palpated per rectum at 10-d intervals after 1 to 2 month postpartum. Cows (n=11), which showed estrus (Day 0) within 5 d of the PGF(2 alpha), and had a CL on Day 7, received ET. Cows in the Ovsynch+CIDR group (n=19) underwent the Ovsynch protocol plus a CIDR for 7 d (GnRH analogue and CIDR on Day-9, PGF(2alpha) analogue with CIDR removal on Day-2, and GnRH analogue on Day 0), with ET on Day 7. The ovulation synchronization (100%) and embryo transfer (100%) rates in the Ovsynch+CIDR group were greater (P<0.01) than the estrus synchronization (66.7%) and the embryo transfer (61.1%) rates in the PGF group. The postpartum interval at ET in the Ovsynch+CIDR group (62.5 +/- 2.5 d) was shorter (P<0.01) than in the PGF group (74.9 +/- 3.9 d). The pregnancy rate in the Ovsynch+CIDR group (57.9%) did not differ significantly from that in the PGF group (50.0%). Plasma progesterone concentrations were not significantly different in the two groups on Days 0, 1, 2, 5, 7, 14 and 21. In summary, higher synchronization and transfer rates, and shorter postpartum interval to ET, can be achieved with timed ET following the Ovsynch plus CIDR protocol than after estrus with the single PGF(2 alpha) treatment followed by ET in suckled postpartum recipient beef cows. Pregnancy rates were similar. Also, the increase in blood progesterone concentrations during luteal development following ovulation synchronized by the Ovsynch plus CIDR protocol was similar to that after estrus induced by the PGF(2 alpha) treatment.     

Some Measurement Results on Sulfate Aerosol Concentration in and above a Pine Canopy

Profile of sulfate aerosol (SO₄ ^2?) concentration was measured for four days at six heights in and above a 15m-high canopy of pine plantation during 6th and 10th August. 1999. The concentration was the lowest (about 2 nmol/m³) on 6th, and gradually increased to 9th showing the maximum values of about 13 nmol/m³, and then decreased to 2 nmol/m³ on 10th. The vertical profiles of SO₄ ^2? concentration showed mostly higher in the canopy than above the canopy. As for the vertical profiles above the canopy on 8th and 10th, the minimum was observed just above the canopy (16m), showing SO₄ ^2? transport from the upper air layer to the canopy. While on 9th the profiles that are higher concentration just above the canopy and lower at the upper air layer were observed, suggesting SO₄ ^2? emission from the canopy to the upper air layer.     

Effect of Simulated Acid Rain on Deterioration of Concrete

In the study the long-term exposure tests to simulated acid rain were performed in order to clarify the effect of acid rain on deterioration of concrete. Mortar specimens with 40 mm in width, 15 mm in thickness and 160 mm in length were used for the tests. At each time after the fixed rainfall was attained, those were tested physically and analyzed chemically. Finally total rainfall of 9000mm was given to the specimens. From the test results, it was confirmed that the eroded depth of the specimen has a good linear relation to the total rainfall under simulated acid rain with various pH. Surface erosion rates of the mortar specimens with an ordinary mix proportion under simulated acid rains with pH 3.0 and 2.5 were about 1.2 and three times larger than that under pH 5.6, respectively. It was also confirmed that flexural strength of the specimens with an ordinary mix proportion hardly changed under low pH simulated acid rain even after total rainfall of 9000mm was given.     

Oligotrophic bacteria on organic debris and plant roots in a paddy field soil

The distribution of heterotrophic bacteria on organic debris and roots of rice plants in a paddy field were studied. The heterotrophic bacteria consisted of two main groups: those which grew on full-strength nutrient broth (NB) and those which did not grow on NB but on a 100-fold dilution of NB (DNB). The latter group was called ‘DNB organisms’ and were considered to be oligotrophic. In both manured and unmanured soils, DNB organisms were predominant in the bacterial communities on organic debris and the rice roots throughout most of the entire period of rice cultivation, although a transient decrease in the proportion of DNB organisms was observed immediately after an application of manure. Morphological and physiological characteristics of DNB isolates from organic debris and rice roots were studied: five types of cell shape were observed, (1) regular rods, (2) filament-forming rods, (3) irregular rods, (4) prosthecate organisms and (5) large oval cells. Regular rods (42% of the total DNB isolates) and irregular rods (46%) were abundant. The ecological roles of DNB organisms in paddy soil are discussed in relation to their physiological characteristics.