Suspected Vestia foetida poisoning in young goats

CASE HISTORY: Two crossbred, castrated male goats, a 5-month-old and an 8-month-old, were observed ingesting Vestia foetida (Solanaceae). Later, the goats were seen standing splay-legged and apparently disoriented.

CLINICAL FINDINGS: When examined, both goats were in sternal recumbency and had mydriasis; the younger goat had a diminished menace response. When the goats were made to stand, they were ataxic and had muscle fasciculations of the hindquarters and face. Both had halitosis consistent with the odour of crushed Vestia leaves. The animals were treated with a mixture of vitamins and intravenous diazepam. The older goat recovered but the younger goat died and was necropsied. This animal had severe periacinar necrosis and fatty change in the liver, as well as fatty nephrosis.

DIAGNOSIS: Probable Vestia foetida poisoning.

CLINICAL RELEVANCE: The introduction of Vestia foetida to New Zealand and the apparent palatability of the plant necessitate that veterinarians and owners be knowledgeable about its potential toxicity. Differential diagnoses for the liver lesions (in New Zealand) would include Cestrum poisoning, acute seneciosis, acute blue-green algal poisoning, and acute and chronic copper poisoning.     

Protocol: Optimised methodology for isolation of nuclei from leaves of species in the <Emphasis Type="Italic">Solanaceae</Emphasis> and <Emphasis Type="Italic">Rosaceae</Emphasis> families

In this study, a protocol is described for rapid preparation of an enriched, reasonably pure fraction of nuclear proteins from the leaves of tobacco (Nicotiana tabacum), potato (Solanum tuberosum) and apple (Malus domestica). The protocol gives reproducible results and can be carried out quickly in 2 hours. Tissue extracts clarified with filtration were treated with non-ionic detergent (Triton X-100) to lyse membranes of contaminating organelles. Nuclei were collected from a 60% Percoll layer of density gradient following low-speed centrifugation. Western blot analysis using antibodies to marker proteins of organelles indicated that the nuclear protein fractions were highly enriched and free or nearly free of proteins from the endoplasmic reticulum and chloroplasts.     

Occurrence and survival of potato scab pathogens (Streptomyces species) on tuber lesions: quick diagnosis based on a PCR-based assay

A time-saving and cost-effective polymerase chain reaction (PCR)-based method was developed for species-specific detection of the scab pathogens ( Streptomyces scabies and S. turgidiscabies ) prevalent in potato ( Solanum tuberosum ) in northern Scandinavia. Species specificity of primers was verified using a collection of previously characterized Streptomyces strains isolated from potato scab lesions in Finland and Sweden. A total of 1245 scab lesions was tested from potato cvs Matilda and Sabina grown in the field in two geographic regions of Finland in 2000 and 2001. Freshly harvested or stored potato tubers were incubated at room temperature (18–21°C) under humid conditions for a few days. Bacterial growth was collected from scab lesions for DNA isolation and PCR. The two scab pathogens were detected in the same potato fields, tubers and scab lesions. The relative incidence of S. scabies was high in freshly harvested tubers but was much lower than that of S. turgidiscabies following storage. Both pathogens were seed-transmitted in Matilda and Sabina after 24 weeks of storage at 4°C.     

Phylogenetic Analysis of 16S rRNA Genes and PCR Analysis of the nec1 Gene from Streptomyces spp. Causing Common Scab, Pitted Scab, and Netted Scab in Finland

ABSTRACT The sequences of the 16S rRNA genes (nucleotides 29 to 1,521) from various Streptomyces strains pathogenic to potato were compared. These included 10 pathogenic Streptomyces strains isolated from potato scab lesions in Finland, the type strains of S. aureofaciens NRRL 2209(T) and S. lydicus ATCC 25470(T), 'S. griseus subsp. scabies' ATCC 10246, and two S. griseus strains that were originally deposited to the collection as pathogens. The nucleotide sequence (>94.5% sequence identity [SI]) and length (1,469 to 1,481 nucleotides) of the analyzed region varied. Phylogenetic analysis of 16S rRNA genes placed Finnish strains into three species, supported by previously characterized morphological and physiological traits. Six Finnish strains, including two strains that deviated from the others in one trait (no spiral sporophores or D-xylose utilization), had identical 16S rRNA genes and were identified as S. scabies (99.9% SI to S. scabies ATCC 49173). Three Finnish strains were identified as S. turgidiscabies, a species previously described only in Japan (99.9% SI to S. turgidiscabies ATCC 700248). Finnish strain 317 and S. aureofaciens NRRL 2209 (99.8% SI) were placed in a distinct phylogenetic cluster together with Kitosatospora spp., which suggests that S. aureofaciens may belong to the recently revived genus Kitosatospora. In pathogenicity tests, S. scabies caused characteristic symptoms of common scab, S. turgidiscabies caused mainly pitted scab, and S. aureofaciens caused netted scab and necrotic lesions on stolons of potato cultivars Bintje and Matilda in the greenhouse. The nec1 gene and the intergenic region between nec1 and the 5' transposase pseudogene ORFtnp were successfully amplified by polymerase chain reaction from S. scabies ATCC 49173 and the pathogenic Finnish strains of S. scabies, but not from a nonpathogenic strain of S. scabies, three pathogenic and two nonpathogenic strains of S. turgidiscabies, and S. aureofaciens.     

Infection with Rhizoctonia solani induces defense genes and systemic resistance in potato sprouts grown without light

Rhizoctonia solani is an important soilborne and seedborne fungal pathogen of potato (Solanum tuberosum). The initial infection of sprouts prior to emergence causes lesions and may be lethal to the sprout or sprout tip, which results in initiation and compensatory growth of new sprouts. They emerge successfully and do not suffer significant damage. The mechanism behind this recovery phenomenon is not known. It was hypothesized that infection may induce pathogen defense in sprouts, which was investigated in the present study. Tubers were sprouted in cool and moist conditions in darkness to mimic conditions beneath soil. The basal portion of the sprout was isolated from the apical portion with a soft plastic collar and inoculated with highly virulent R. solani. Induction of defense-related responses was monitored in the apical portion using microarray and quantitative polymerase chain reaction techniques at 48 and 120 h postinoculation (hpi) and by challenge-inoculation with R. solani in two experiments. Differential expression of 122 and 779 genes, including many well-characterized defense-related genes, was detected at 48 and 120 hpi, respectively. The apical portion of the sprout also expressed resistance which inhibited secondary infection of the sprouts. The observed systemic induction of resistance in sprouts upon infection with virulent R. solani provides novel information about pathogen defense in potato before the plant emerges and becomes photosynthetically active. These results advance our understanding of the little studied subject of pathogen defense in subterranean parts of plants.     

Evaluation of a diagnostic microarray for the detection of major bacterial pathogens of potato from tuber samples

Potato can be infected with many bacterial pathogens, the detection of which is necessary in seed certification. In this study, a diagnostic microarray previously tested for specificity of probes for detecting the potato bacteria causing blackleg and soft rot (Pectobacterium atrosepticum, Pectobacterium carotovorum, and Dickeya spp.), ring rot (Clavibacter. michiganensis subsp. sepedonicus), scab (Streptomyces scabies and Streptomyces turgidiscabies) and brown rot (Ralstonia solanacearum) from pure culture was evaluated for analytical sensitivity when testing directly from tuber samples. The microarray readily detected all the bacterial species when 100 ng of the target bacterial DNA from pure culture was mixed with DNA from soil microbes and potato. However, detection was inconsistent when total DNA isolated directly from infected tubers or enriched bacterial culture was used. While the high specificity of the probes could be confirmed from the results of the DNA cocktail experiment used as a control, the study demonstrated that the level of analytical sensitivity of the microarray under the tested condition was not sufficient to detect bacteria directly from tubers. Therefore, in addition to the cost and organizational complexities, the low analytical sensitivity and limited reproducibility of the microarray are constraints for establishing the platform for routine detection of potato bacterial pathogens from tuber samples.     

铲式成穴器成穴性能的试验研究

根据铲式成穴器的成穴机理确定了其结构参数和工作参数的设计变化范围并按照二次回归试验设计方法进行了成穴器参数地成穴性能影响的试验,通过对试验结果垢统计分析,查明了成穴器参数对穴孔长度,穴孔宽度穴距和滑移系数的影响。     

Variability of Peronospora Sparsa (syn. P. rubi) in Finland as Measured by Amplified Fragment Length Polymorphism

Downy mildew caused by Peronospora sparsa (syn. P. rubi) is a serious threat to commercial cultivation of arctic bramble (Rubus arcticus subsp. arcticus) in Finland. P. sparsa is distributed throughout the country in cultivated and wild arctic bramble and in cloudberry (R. chamaemorus). A total of 36 isolates of P. sparsa collected from these hosts was analysed for amplified fragment length polymorphism (AFLP). Of the 226 markers scored, 223 were polymorphic and all isolates of P. sparsa had unique AFLP fingerprints, which indicated high levels of genetic variability. An UPGMA clustering analysis of the isolates did not reveal any genetically distinguishable strains. The isolates were grouped neither according to the geographic origin nor the host from which they were isolated. Isolates of P. sparsa obtained from wild arctic bramble and one from cloudberry readily infected the leaves of the cultivated arctic bramble (cultivar Pima). Also, P. sparsa isolated from cultivated arctic bramble infected the leaves of wild arctic bramble. These data suggest that P. sparsa may be disseminated from wild arctic bramble and cloudberry to cultivated arctic bramble in the field.