Isolates of Verticillium dahliae Pathogenic to Crucifers Are of at Least Three Distinct Molecular Types

ABSTRACT Diverse isolates of the soilborne wilt fungi Verticillium dahliae and V. albo-atrum were studied to understand the nature and origins of those infecting cruciferous hosts. All isolates from cruciferous crops produced microsclerotia, and the majority produced long conidia with a high nuclear DNA content; these isolates were divided into two groups by amplified fragment length polymorphism (AFLP) analysis. One group could be subdivided by other criteria such as rRNA sequences and mitochondrial DNA restriction fragment length polymorphism (RFLP) analysis. Two crucifer isolates were short spored and had a low nuclear DNA content. The results are consistent with the crucifer isolates being interspecific hybrids. The long-spored isolates are best regarded as amphihaploids (or allodiploids) with the AFLP groups probably each representing separate interspecific hybridization events. The short-spored crucifer isolates appear to be derived from interspecific hybrids and are here called 'secondary haploids'. Molecular evidence suggests that one parent in the crosses was similar to V. dahliae. The other parent of the amphihaploids seems to have been more similar to V. albo-atrum than to V. dahliae, but was distinct from all isolates of either species so far studied. The implications for the taxonomy of crucifer isolates are discussed and the use of the name V. longisporum, proposed elsewhere for just some of these isolates, is discouraged.     

Genetic diversity and pathogenic variability among isolates of colletotrichum species from strawberry

ABSTRACT Ninety-five isolates of Colletotrichum including 81 isolates of C. acutatum (62 from strawberry) and 14 isolates of C. gloeosporioides (13 from strawberry) were characterized by various molecular methods and pathogenicity tests. Results based on random amplified polymorphic DNA (RAPD) polymorphism and internal transcribed spacer (ITS) 2 sequence data provided clear genetic evidence of two subgroups in C. acutatum. The first subgroup, characterized as CA-clonal, included only isolates from strawberry and exhibited identical RAPD patterns and nearly identical ITS2 sequence analysis. A larger genetic group, CA-variable, included isolates from various hosts and exhibited variable RAPD patterns and divergent ITS2 sequence analysis. Within the C. acutatum population isolated from strawberry, the CA-clonal group is prevalent in Europe (54 isolates of 62). A subset of European C. acutatum isolates isolated from strawberry and representing the CA-clonal and CA-variable groups was assigned to two pathogenicity groups. No correlation could be drawn between genetic and pathogenicity groups. On the basis of molecular data, it is proposed that the CA-clonal subgroup contains closely related, highly virulent C. acutatum isolates that may have developed host specialization to strawberry. C. gloeosporioides isolates from Europe, which were rarely observed were either slightly or nonpathogenic on strawberry. The absence of correlation between genetic polymorphism and geographical origin in Colletotrichum spp. suggests a worldwide dissemination of isolates, probably through international plant exchanges.     

Efficacy of extended pirlimycin hydrochloride therapy for treatment of environmental Streptococcus spp and Staphylococcus aureus intramammary infections in lactating dairy cows

Fifty-one chronically infected lactating dairy cows were used to evaluate the efficacy of extended pirlimycin therapy regimens for treatment of intramammary infections by environmental Streptococcus spp and Staphylococcus aureus. Cows (n = 47) with one or more infected mammary quarters were blocked by parity and randomly allocated to one of three groups for treatment with pirlimycin (50 mg/mammary quarter) as follows: one treatment per day for 2 days (n = 36 infected mammary quarters); one treatment per day for 5 days (n = 36 infected mammary quarters); and one treatment per day for 8 days (n = 20 infected mammary quarters). Four cows with nine infected mammary quarters were included as untreated controls. Milk samples from each mammary quarter were collected 7 days before treatment, immediately before treatment, and weekly for 4 weeks after the final treatment for microbiological evaluation. A bacteriologic cure was defined as a treated, infected quarter that was bacteriologically negative for the presence of previously identified bacteria at weekly intervals after treatment. Efficacy of pirlimycin therapy against intramammary infections caused by environmental Streptococcus spp and S. aureus was 44.4%, 61.1%, and 95.0% for the 2-, 5-, and 8-day treatment regimens, respectively. None of the infections in the untreated control quarters was cured. Significant differences in efficacy were detected between all pirlimycin groups and the untreated control group, between the 8- and 2-day treatment regimens, and between the 8-day and 5-day treatment regimens (P < or = .05). Results of this study indicate that extended pirlimycin therapy was effective in eliminating intramammary infections caused by environmental streptococci and S. aureus in lactating dairy cows.     

Matrix metalloproteinase activity in tumor, stromal tissue, and serum from cats with malignancies

Matrix metalloproteinases (MMPs) are enzymes that play key roles in angiogenesis, tumor invasion, and metastasis in a wide variety of species. The purpose of this study was to evaluate pro and active MMP 2 and 9 concentrations in tumor, normal stromal tissue, and serum from tumor-bearing cats. We hypothesized that serum concentrations of pro and active forms of MMPs 2 and 9 would be predictive of MMP concentrations in tumor tissue and that these MMP concentrations would correlate with the histopathologic grade of the malignancies. Pro and active forms of MMPs 2 and 9 were determined by gelatin zymography and subsequent computerized densitometry from tumor and nearby stromal tissue and serum from 49 cats with various malignancies. The serum concentrations of MMPs from these tumor-bearing cats were compared with serum concentrations of MMPs from 44 normal cats of similar age and gender. Measurable concentrations of MMPs 2 and 9 were found within tumor, stromal, and serum samples. Mean concentrations of total pro and active MMPs 2 and 9 within tumor tissue were significantly higher (P values <.0001, .0031, <.001, and .0064, respectively) when compared with stromal tissue from the same animals. Serum MMP concentrations from tumor-bearing cats were higher than those from normal cats. Poor correlation was found between serum MMP concentrations and tissue MMP concentrations of increasing histologic grades of malignancies.     

Agreement among surgical pathologists evaluating routine histologic sections of digits amputated from cats and dogs.

Agreement among pathologists interpreting histologic specimens is an area of interest within human pathology, but little work in this area has been reported in the veterinary literature. Agreement among pathologists evaluating routine histologic sections of amputated digits from cats and dogs submitted to multiple diagnostic centers was examined. Histologic sections from surgical specimens were reviewed in a blinded fashion by two pathologists, and a comparison to the original diagnosis, as stated in the diagnostic report, was recorded. A total of 513 cases were reviewed, and complete agreement was reached in 409 (79.7%). Of the 104 instances of disagreement, 77 (74.0%) were considered to be of clinical significance. The diagnosis of keratoacanthoma was disagreed with in 19 of 21 diagnoses (90.4%). No other individual diagnosis was similarly disputed. The overall level of disagreement is large and is similar to that reported in human pathology and suggests that further study of this issue would be useful in veterinary pathology.     

Major gene resistance in Brassica napus (oilseed rape) is overcome by changes in virulence of populations of Leptosphaeria maculans in France and Australia

Resistance of Brassica napus (oilseed rape, canola) conferred by three different major resistance genes has been overcome by changes in virulence of Leptosphaeria maculans populations in France and Australia. In South Australia where B. napus cultivars with major gene resistance derived from Brassica rapa ssp. sylvestris were grown extensively, resistance was rendered ineffective within 3 years of commercial release of the cultivar. Disease severity was higher on cultivars with sylvestris-derived resistance than cultivars with polygenic resistance. This Australian situation is compared to that in France, where resistance conferred by the Rlm1 gene was overcome nation-wide in 5 years under commercial cropping practices, and also where a source of resistance introgressed into B. napus from B. juncea was rendered inefficient in 3 years in experimental field plots near Rennes.     

A Combination Chemotherapy Protocol (VELCAP-L) for Dogs with Lymphoma

Ninety-eight dogs with lymphoma treated with a 5-drug combination chemotherapy regimen (vincristine, L-asparaginase. cyclophosphamide, doxorubicin, prednisone [VELCAP-L]) were evaluated for pretreatment characteristics predictive for response and remission duration. The complete remission rate was 69%, with a median remission duration of 55 weeks. Dogs with advanced stage of disease, constitutional signs, dogs that were older, and dogs that were dyspneic were less likely to achieve remission. Once in remission, small dogs and dogs without pretreatment thrombocytopenia were likely to have longer remission duration. Toxicoses were frequent, but rarely fatal, and no predictitive factors were found for a dog developing toxicoses. VELCAP-L is an effective treatment for dogs in stage I-III lymphoma, particularly in young, small animals.     

First report of Cryptosporidium deer-like genotype in Kenyan cattle

The objective of the study was to identify Cryptosporidium genotypes from feces collected from urban and peri-urban dairy cattle in Nairobi, Kenya, in order to determine their zoonotic potential. DNA was extracted from 34 samples that were diagnosed positive by the modified Ziehl-Neelsen technique. Two Cryptosporidium isolates examined at the 18S rRNA locus were identified as the deer-like genotype by DNA sequencing. As public health officials are facing the difficult decision whether to allow urban livestock production because of its economic benefits and a livelihood asset to the urban communities, or to ban it for its public health risks, the finding of non-zoonotic genotypes in a smallholder dairy system has significant public health as well as economic implications that merit further investigation.     

Plasma volume and electrolyte changes following intravenous infusion of hypertonic hydroxyethyl starch versus mannitol in healthy dogs

In a prospective cross-over study, the duration and magnitude of effect on the electrolyte and plasma volume changes of intravenous (IV) hypertonic hydroxyethyl starch (hyperHES) (7.5%/6%) and mannitol (20%) were compared. Eight Beagle dogs received an IV infusion of 4mL/kg hyperHES (group H) and 4mL/kg mannitol 20% (group M) on separate occasions. Urine and blood samples were taken in the first (T(60)) and second (T(120)) hour after infusion. Significant increases in plasma volume at each time point in group H and M were noted when compared to baseline (start of infusion=T(0)) level. There was no significant difference between groups. Both fluids resulted in diuresis, although no significant difference between groups was noted. A significant increase in plasma sodium (Na) was demonstrated in group H between T(0) and T(60) with a significant increase in the Na and chloride (Cl) fractional excretion (FE) between T(0), T(60) and T(120). In group M no changes in plasma electrolyte concentrations were detected, although FE of Na, Cl and K was increased significantly between T(0) and T(60). In conclusion, hyperHES and mannitol appear to have a volume expanding effect lasting at least 120 min. The hypernatraemia induced by hyperHES was minimal compared to previous reports of hypertonic saline use, and no clinical side effects were noted. HyperHES showed comparable effects to mannitol in increasing plasma volume and diuresis and could be considered for these applications.