Identification of Mulberry Dwarf Phytoplasmas in the Genital Organs and Eggs of Leafhopper Hishimonoides sellatiformis

ABSTRACT The presence of mulberry dwarf (MD) phytoplasmas in organs of the inoculative vector insects Hishimonoides sellatiformis and Hishimonus sellatus was determined by means of electron microscopy (EM) and polymerase chain reaction (PCR) assays. Many MD phytoplasmas were detected in genital organs as well as in the intestines, salivary glands, brains, fat bodies, and thoracic ganglia of Hishimonoides sellatiformis, but only in the intestine and salivary glands of Hishimonus sellatus. Many phytoplasmas with characteristic morphology were observed via EM in ovaries, seminal receptacles, and testes, and they were further identified by PCR assays with group I-specific primers. In addition, the organisms were detected by direct or nested PCR assays in eggs (head pigmentation stage of embryos) laid on mulberry shoots by inoculative leafhoppers and in the newly hatched nymphs from these eggs. These findings indicate that transovarial transmission of MD phytoplasmas occurs in Hishimonoides sellatiformis.     

Frequency of enteropathogen detection in suckling and weaned pigs with diarrhea in Japan.

Fecal samples from suckling (n=153) and weaned (n=116) piglets with diarrhea in Japan were examined for shedding of viral, bacterial, and parasitic pathogens using culture, microscopic, and polymerase chain reaction methods. In suckling piglets, diarrhea was attributed to infection with a single etiologic agent in 60.8% of cases and with combinations of agents in 22.2%. In weaned piglets, diarrhea was attributed to a single etiologic agent in 43.1% and to combinations of agents in 47.4% of cases. Rotavirus was the most prevalent agent in suckling (67.3%) and weaned (65.5%) piglets. The detection of other pathogens was associated with age of the animals examined. Coccidia were predominantly isolated from suckling piglets, whereas Escherichia coli was found predominantly in weaned piglets. Although a relationship was not observed between detection rate of rotavirus and age of piglets, a single group of rotavirus was detected in 87.5% of suckling piglets whereas multiple groups were detected in 51.6% of weaned piglets. The results of this study confirm that diarrhea in piglets can, to a variable degree, be causally associated with multiple agents. Additionally, these results suggest reasons why this syndrome can be difficult to control.     

Epidemiological investigation of the prevalence and features of postweaning multisystemic wasting syndrome in Japan.

To investigate the prevalence and features of postweaning multisystemic wasting syndrome (PMWS) in Japan, an epidemiological study was conducted in 692 weaned pigs with various clinical signs, commonly including wasting or weight loss, collected from 129 swine farms between 2000 and 2003. The presence of PMWS was diagnosed by the detection of characteristic histological lesions and moderate to large amounts of porcine circovirus type 2 (PCV2) antigen within the lesions in multiple lymphoid tissues. Postweaning multisystemic wasting syndrome was positive in 23.4% of pigs (162/692) over the course of the study, and occurred in 50.4% of the farms (65/129). Mortality in 30-120-day-old pigs in the farms positive for PMWS varied from 0.1 to 32.0%. No significant difference in mortality was seen between PMWS-positive and -negative farms (P = 0.1). However, mortality was significantly higher in the PMWS-positive farms where PMWS was diagnosed in more than 50% of the pigs examined compared to farms negative for PMWS (P = 0.02). These findings indicate that PMWS has spread widely in Japan. Moreover it may exist in variable forms in swine farms, including an epidemic form or a subtle endemic or sporadic form. A case-control study suggested that risk factors for the occurrence of PMWS include porcine reproductive and respiratory syndrome (PRRS) pneumonias and Mycoplasma hyorhinis infection.     

Identification of Petunia hybrida cultivars that diurnally emit floral fragrances

In order to identify genetic resources for breeding fragrant petunias for use as bedding plants, volatile compounds released by day from the flowers of 40 commercial Petunia hybrida cultivars were analyzed using a solid-phase micro-extraction technique coupled with GC–MS. The three cultivars with solid deep-blue flowers that accumulate malvidin in corollas with high tissue pH were found to emit abundant iso-eugenol as the principal floral fragrance. Several other cultivars that emitted considerable amounts of methylbenzoate and/or benzylbenzoate from the flower were also identified. Association between the floral fragrance and the other floral traits such as floral anthocyanin composition and corolla-tissue pH was discussed.     

Landscape improvement, or ongoing degradation – reconciling apparent contradictions from the arid rangelands of Western Australia

Recent quantitative site-based monitoring and qualitative aerial and ground traverses provide contrasting assessments of the health of much of the arid shrublands of Western Australia extensively grazed by livestock (‘rangelands’). Although these results seem incompatible, we explain the apparent contradictions based on landscape succession processes operating at multiple levels of ecological organisation. Specifically, we suggest that the intact areas in which site-based monitoring is conducted are contracting as catchment canalisation and desiccation increase. However, the impacts of these processes have not yet become manifest at the site scale. The site-based system addresses important regional questions. These relate to the large, relatively intact areas away from most active surface flows, which should be a focus for resource conservation, given practical limits to repairing widespread degradation with low management inputs. We provide a complementary set of questions to provide a more comprehensive audit of rangeland dynamics in the context of underlying hierarchical landscape patterns and processes that might threaten intact areas. We recognise the need to match questions and levels of ecological organisation and the implications these have for sampling. We also recognise the difficulty in producing concise statements of change for clients when reporting on complex ecological issues and processes. Without a clearly articulated, and well understood, hierarchical model of pattern and process within which apparently contradictory findings can be reported meaningfully, policy makers may be confused by the results, with the consequent risk of policy inaction.     

Mottle necrosis of sweet potato caused by <Emphasis Type="Italic">Pythium scleroteichum</Emphasis> in Japan and varietal difference in susceptibility to the disease

Severe mottle necrosis (Shirogusare-byo in Japanese) was found on mature tubers of sweet potato (Ipomoea batatas) in Ibaraki Prefecture in October 2004. The causal organism was identified as Pythium scleroteichum hitherto unknown in Japan. Sweet potato cultivar Purple Sweet Lord was more susceptible than cultivars Beniazuma, Benimasari, Koukei-14, and Tamayutaka to the pathogen at 25°C, while this difference in the susceptibility was not clear at 15°C.     

A Conjugative Plasmid Carrying the efe Gene for the Ethylene-Forming Enzyme Isolated from Pseudomonas syringae pv. glycinea

ABSTRACT Previous work suggested that the efe gene encoding the ethylene-forming enzyme was present in the plasmids of three pathovars of Pseudomonas syringae including glycinea, phaseolicola (kudzu strains), and cannabina. However, no direct evidence to support this assumption had been presented. In the current study, we isolated the conjugative plasmid harboring the efe gene (ethylene plasmid) designated pETH2 from P. syringae pv. glycinea MAFF301683. pETH2 was detected by Southern blot hybridization using the efe probe, marked with the transposon mini-Tn5-Km1, and transferred into P. syringae Ni27(n), which does not produce ethylene. The transconjugant Ni27(n) (pETH2) produced ethylene at a level similar to pv. glycinea MAFF301683. In addition, the plasmid designated pCOR2, which encodes coronatine biosynthesis genes, was detected in the same strain. Although the molecular size of the plasmid pCOR2 was not easily distinguishable from pETH2, pCOR2 transferred independently into Ni27(n) and the transconjugants produced coronatine. These findings suggested that the efe gene has been horizontally dispersed among pathovars of P. syringae by plasmid-mediated conjugation in nature.     

Diazepam potentiates the positive inotropic effects of histamine and forskolin in guinea-pig papillary muscles

There have been diverse reports on the effects of diazepam on cardiac contractility. The purpose of this study was to examine whether diazepam modifies the inotropic response elicited by histamine on an isolated guinea-pig papillary muscle. The responses of electrically driven papillary muscle to histamine and cyclic AMP-related inotropic agents were recorded in the absence and in the presence of diazepam. Histamine and forskolin, which directly stimulate adenylate cyclase, significantly increased the contractile force in the papillary muscle in a concentration-dependent manner. A histaminergic H₂-receptor antagonist, cimetidine, but not a H₁-receptor antagonist, diphenhydramine, at 10 μ M produced a rightward shift in the concentration-response curve for histamine. Diazepam (10 μ M ) shifted the concentration-response curve for histamine and forskolin to the left by 1.8 and 1.6 times, respectively. Neither a central type (fulmazenil) nor a peripheral type (PK11195) of benzodiazepine receptor antagonist modified the effect of diazepam on the histaminergic-evoked contraction. Phosphodiesterase blockade by 3-isobutyl-1-methylxanthine shifted the concentration-dependent curve for histamine to the left. A combination of 3-isobutyl-1-methylxanthine also produced a leftward shift of the curve. However, there was no significant difference between the 3-isobutyl-1-methylxanthine only group and the combination group. These results indicate that diazepam potentiates the positive inotropic effect produced by histamine, probably mediated via an increase in cyclic AMP levels induced by histamine.     

Intracellular recording of rat neuron activity at the medial preoptic area of the hypothalamus using triangular wave microelectrode oscillation

A triangular wave microelectrode oscillating apparatus was constructed to evaluate an intracellular recording of rat neuron activity in the medial preoptic area (POA) of the hypothalamus. In this apparatus, electrodes passed a current with a frequency of 1.0 to 1.8 kHz and a voltage of 2.2 to 3.2 V and produced micro-oscillation of the electrode tip. The electrode was inserted into a neuron of the rat POA in vivo. In vivo recording of the activity of the rat POA neuron was possible. By means of electrical stimulation of the median eminence arcuate of the hypothalamus, an intracellular recording of antidromic, orthodromic or non-responding neuron was also possible. As a result, various components of the action potential such as the resting, threshold and spike potentials, and depolarization and repolarization such as after-hyperpolarization and after-depolarization were observed. The resting potentials ranged from 45 to 90 mV, and POA neurons possessed action potentials of almost the same magnitude. Several problems, however, remain to be solved. In general, the time available for the intracellular recordings is too short. The cells survive only for 15 minutes at the longest and may die in only a few minutes. An improvement of the apparatus was mandatory.