Phylogenetic analysis of cercospora and mycosphaerella based on the internal transcribed spacer region of ribosomal DNA

ABSTRACT Most of the 3,000 named species in the genus Cercospora have no known sexual stage, although a Mycosphaerella teleomorph has been identified for a few. Mycosphaerella is an extremely large and important genus of plant pathogens, with more than 1,800 named species and at least 43 associated anamorph genera. The goal of this research was to perform a large-scale phylogenetic analysis to test hypotheses about the past evolutionary history of Cercospora and Mycosphaerella. Based on the phylogenetic analysis of internal transcribed spacer (ITS) sequence data (ITS1, 5.8S rRNA gene, ITS2), the genus Mycosphaerella is monophyletic. In contrast, many anamorph genera within Mycosphaerella were polyphyletic and were not useful for grouping species. One exception was Cercospora, which formed a highly supported monophyletic group. Most Cercospora species from cereal crops formed a subgroup within the main Cercospora cluster. Only species within the Cercospora cluster produced the toxin cercosporin, suggesting that the ability to produce this compound had a single evolutionary origin. Intraspecific variation for 25 taxa in the Mycosphaerella clade averaged 1.7 nucleotides (nts) in the ITS region. Thus, isolates with ITS sequences that differ by two or more nucleotides may be distinct species. ITS sequences of groups I and II of the gray leaf spot pathogen Cercospora zeae-maydis differed by 7 nts and clearly represent different species. There were 6.5 nt differences on average between the ITS sequences of the sorghum pathogen Cercospora sorghi and the maize pathogen Cercospora sorghi var. maydis, indicating that the latter is a separate species and not simply a variety of Cercospora sorghi. The large monophyletic Mycosphaerella cluster contained a number of anamorph genera with no known teleomorph associations. Therefore, the number of anamorph genera related to Mycosphaerella may be much larger than suspected previously.     

Strand-specific postreplicative processing of mammalian telomeres

Telomeres are specialized nucleoprotein structures that stabilize the ends of linear eukaryotic chromosomes. In mammalian cells, abrogation of telomeric repeat binding factor TRF2 or DNA-dependent protein kinase (DNA-PK) activity causes end-to-end chromosomal fusion, thus establishing an essential role for these proteins in telomere function. Here we show that TRF2-mediated end-capping occurs after telomere replication. The postreplicative requirement for TRF2 and DNA-PKcs, the catalytic subunit of DNA-PK, is confined to only half of the telomeres, namely, those that were produced by leading-strand DNA synthesis. These results demonstrate a crucial difference in postreplicative processing of telomeres that is linked to their mode of replication.     

Detection of koi herpesvirus DNA in tissues of infected fish

A newly recognized herpesvirus, koi herpesvirus or KHV, causes a lethal disease in common carp, Cyprinus carpio , and its colourful strain known as koi or fancy carp. In this study, we report new outbreaks of the disease, present initial characterization of the KHV genome, and describe assays for detection of KHV DNA in infected cells and tissues of infected fish. Restriction endonuclease (RE) profiles of viral DNA derived from two epidemiologically distinct KHV isolates were identical to each other. Cloned KHV BamHI and SphI DNA probes specifically hybridized to KHV DNA, but not to DNAs derived from a variety of other fish herpesviruses. The KHV DNA probes detected KHV DNA in tissues of experimentally infected koi fish by DNA hybridization. The KHV specific polymerase chain assays (PCR) were developed for rapid detection and confirmation of KHV DNA in tissues of infected fish.     

Identification of trout and salmon bloods by simple immunological technique and by electrofocusing patterns of red cell enzyme superoxide dismutase

Species identification of animal bloods is readily achieved by immunological tests. Differentiation among fish species on this basis is more difficult although considerable success has been achieved on the basis of both inter- and intra-specific differences in their serum proteins. This report describes a method for the identification of the different species of fish within the Salmonidae family and some coarse fish families on the basis of an immunological test and electrofocusing patterns of the enzyme superoxide dismutase from the red cell. The immunological technique relies on the development of a specific anti-trout (Salmonidae) serum which is used initially to differentiate the blood of a Salmonidae from other freshwater fish. Further discrimination, within the Salmonidae, is made on the basis of the different polymorphic forms of the enzyme superoxide dismutase separated in a pH 2.5 to 8 gradient. Using this technique, it is possible to differentiate among salmon, sea/brown trout, char, cheetah trout, and a number of varieties of rainbow trout.     

Arterial Blood Gases in Dogs with Bacterial Pneumonia

The purpose of this study is to report the aterial blood gas findings in dogs with bacterial pneumonia. Arterial blood gas samples were collected from 62 dogs with culture-confirmed bacterial pneumonia. These results were compared with 46 normal dog arterial blood gas samples. Results demonstrated that respiratory acidosis was not a problem in dogs with pneumonia in this study. Significant evidence of hypoxemia was noted with abnormal mean values in PaO₂ (P<0.001) and the Alveolar-arterial (A-α) gradient (P<0.001).     

Intestinal cryptosporidiosis in chickens

In a retrospective examination of histopathology reports from Aug. 1, 1985, through Sept. 31, 1987, 10 cases of small- or large-intestinal cryptosporidiosis (not epithelial cryptosporidiosis of the bursa of Fabricius) were found in chickens. Infection was evenly distributed among young chickens. Incidence of intestinal cryptosporidiosis increased during 1987. Although all infected birds were clinically ill, signs or gross lesions of intestinal disease were not always present. In all cases, mild to marked histologic lesions were associated with Cryptosporidium sp.; however, intestinal tracts were not cultured for other infectious agents. The numbers of Cryptosporidium sp. and character of inflammatory response were not significantly correlated. A difference (P = 0.01) among intestinal segment (small vs. large) infection with Cryptosporidium was seen. Light-microscopic appearance and organ distribution of Cryptosporidium sp. suggest that in addition to C. baileyi, other Cryptosporidium species infect chickens. Until the diagnostic procedure for outbreaks of gastrointestinal disease in poultry routinely includes histopathology, fecal flotation, and virus, bacteria, and chlamydia cultures, and until species of Cryptosporidium are isolated, identified, reported, and investigated experimentally, the importance of intestinal cryptosporidiosis in chickens will remain unknown.     

Clinical chemistry reference values in two breeds of swine and their changes during percutaneous exposure to soman

Clinical chemistry reference values in blood from 48 nonfasting Chester White/Yorkshire and 48 Hanford Miniature swine were determined. Subsequently, 40 animals of each breed were restrained in a cloth sling and fasted for 24 hours while exposed percutaneously to pinacolyl methylphosphonofluoridate (soman). The range of dosages for the Hanford Miniature swine was 2.0 to 15.8 mg/kg, and for the Chester White/Yorkshire swine, the range was 4.0 to 25.0 mg/kg. Sham-exposed groups, consisting of 8 animals of each breed, were treated in an identical manner, except no anticholinesterase agent was administered. Samples of blood were drawn at 1, 7, 14, and 28 days after soman or sham exposure. In the sham-exposed groups, significant changes from the reference values were observed as a result of the 24-hour restraint. In both breeds, skeletal muscle enzyme activities were increased, plasma cholinesterase activity (ChEPL) was decreased, calcium concentration was decreased, and phosphorus concentration was increased. Percutaneous exposure to soman resulted in decreases of ChEPL and erythrocyte cholinesterase activities (ChERBC). The ChEPL recovered more quickly than the ChERBC in both breeds. Even in asymptomatic swine, the decrease of ChERBC was greater than 60% after 24 hours. In the swine of each breed given the largest dosage, hyperglycemia was apparent in blood samples taken at the onset of apnea, especially when the animal survived for greater than 2 hours. We conclude that both breeds of swine, on the basis of dispersion in clinical chemistry reference values, were equally suited for this type of dermatotoxicity study. The sling method of restraint, however, caused some undesirable changes in biochemical values.