Recent observations on leafy gall in Liliaceae and some other families

Corynebacterium fascians, which causes leafy gall, has been shown to be responsible for the unusual symptoms recently found in lilies. The symptoms are described and the different bacterial isolates compared by biochemical, serological and pathogenicity experiments. No evidence could be found for the existence of specialized strains even though considerable variation in virulence could be demonstrated. Although the role of variation in susceptibility of the different lily cultivars should not be underestimated, it would appear that high inoculum levels ofC. fascians in the soil may be largely responsible for these outbreaks.Also included in this study are the results of biochemical, serological and pathogenicity experiments ofC. fascians which has been isolated fromKalanchoe, Euphorbia, Brodiaea, Hebe andVerbascum.Samenvatting Reeds een aantal jaren was in lelies een woekering in stengelbollen bekend, die echter sporadisch voorkwam en economisch van weinig betekenis was. Tijdens de zomer van 1977 echter trad de ziekte op één enkel perceel zo ernstig op dat zeer aanzienlijke oogstderving het gevolg was. Behalve de stengelbol kan ook de hoofdbol misvormd zijn. Bovendien kunnen misvormde stengelbollen lange spruiten vormen, zgn. sabeltandvorming. Soms is het wortelstelsel gereduceerd; in de kas zijn verdikte stengels waargenomen. De ziekte staat nu bekend onder de naam woekerziekte.Door de PD te Wageningen en het LBO te Lisse werd de bacterieCorynebacterium fascians (Tilford) Dows. uit het zieke materiaal geïsoleerd. In gezamenlijk onderzoek werd nagegaan of de bacterie de veroorzaker is van bovengenoemde symptomen en of hier sprake is van een op lelie gespecialiseerde stam.Biochemische en serologische vergelijkingen tussen isolaten vanC. fascians uit lelie,Kalanchoe, Euphorbia, Brodiaeea, Hebe enVerbascum laten zien dat er zeer weinig verschillen bestaan. Alle isolaten reageren op een standaard antiserum met titers variërend tussen 1: 640–1: 5120.Een oriënterende inoculatieproef werd uitgevoerd op het LBO in 1977. Daarna werden op de PD 10 van de 25 isolaten uit lelie en 8 uit andere waardplanten getoetst op lelie, chrysant en lathyrus. Uit de inoculatieproeven blijkt, dat de bacterie de veroorzaker is van de waargenomen symptomen. Tussen de isolaten blijkt een groot verschil in pathogeniteit te bestaan (12–100% aantasting), zonder dat dit aan herkomst gebonden is. Het voorkomen van op lelie of andere waardplanten gespecialiseerde stammen is daarom niet aan te nemen.In hoeverre de hevige aantasting een gevolg is van een opbouw van de bacterie-populatie door een te nauwe vruchtwisseling en mechanische rooimethoden moet worden nagegaan.     

Molecular and symptom analyses of phytoplasma strains from lettuce reveal a diverse population

ABSTRACT Epidemics of aster yellows in lettuce in Ohio are caused by at least seven distinct phytoplasma strains in the aster yellows (AY) group. Five of the strains are newly reported: AY-BW, AY-WB, AY-BD3, AY-SS, and AY-SG. All seven strains were characterized based on symptoms in aster and lettuce, and by polymerase chain reaction (PCR). Strain AY-BD2 (formerly 'Bolt') causes yellowing and leaf distortion in lettuce and bolting in aster, whereas strain AY-S (formerly 'Severe') causes stunting, leaf clustering, and phyllody. Strain AY-WB causes yellowing and wilting in lettuce and witches'-broom in aster. Strain AY-SG induces horizontal growth in lettuce and aster plants. Strain AY-BW causes chlorosis of emerging leaves and abnormally upright growth of leaf petioles. AY-SS causes symptoms similar to those caused by AY-S but has a different PCR-restriction fragment length polymorphism (RFLP) banding pattern. Strains AY-BD2 and AY-BD-3 cause mild leaf and stem distortion in lettuce but are differentiated by PCR-RFLP. All phytoplasma strains collected from lettuce in Ohio belong to the 16SrI group. AY-WB belongs to the 16SrI-A subgroup and the other six belong to the 16SrI-B subgroup. Five of the seven strains were distinguished from each other by primer typing. The results of phylogenetic analyses of sequences of the 16S rRNA genes were basically consistent with the classification based on PCR-RFLP, in which AY-WB clustered with phytoplasmas of the 16rIA subgroup and the other Ohio lettuce strains clustered with phytoplasmas in the 16SrI-B subgroup.     

Proliferative renal myxosporidiosis in spawning coho salmon (Oncorhynchus kisutch) in British Columbia and Washington

An unidentified myxosporean parasite (CKX) is described from the kidney of approximately 80% of spawning coho salmon Oncorhynchus kisutch (Walbaum) in British Columbia, Canada and Washington, United States of America. Morphological features were described using light and electron microscopy. Sequencing of polymerase chain reaction (PCR) amplified 18S ribosomal RNA gene and in situ hybridisation were used to further characterise CKX. The parasite occurred with a focal distribution within tubule epithelial cells, the tubule lumen and the interstitium as primary cells containing from one to at least 16 secondary cells. Luminal stages were degenerate and sporogony was not observed. In situ hybridisation using a digoxygenin-labelled DNA probe confirmed CKX to be the source of DNA used in PCR analyses. CKX 18S rDNA sequences were most similar (97%) to those of Sphaerospora oncorhynchi. Phylogenetic analysis revealed similarities among the 18S rDNA sequences of CKX, S. oncorhynchi and Myxidium lieberkuehni. CKX is hypothesised to be the abortive extrasporogonic development of a Sphaerospora sp. or Myxidium sp. occurring in immune-incompetent spawning and senescent salmon.     

Rete testis mucinous adenocarcinoma in a dog

A bilateral testicular neoplasm from an 11-year-old mixed-breed male dog was removed surgically and examined histologically. The neoplasm was nonencapsulated and composed of acinar and tubular structures lined by one or more layers of neoplastic polyhedral epithelial cells with an abundant mucinous secretion. On histochemistry, all neoplastic cells and associated secretions were periodic acid-Schiff positive. Some neoplastic cells and all associated secretions were positive on mucicarmine stain, and some neoplastic cells, all the stroma, and associated secretions were positive on alcian blue stain. On immunohistochemistry, the neoplastic cells had strong diffuse cytoplasmic immunoreactivity for cytokeratin and vimentin, weak scattered cytoplasmic immunoreactivity for carcinoembryonic antigen and neuron-specific enolase, and no immunoreactivity for S-100. On the basis of histopathology, histochemistry, and immunohistochemical findings, a diagnosis of mucinous adenocarcinoma of rete testis was made. Rete testis adenocarcinoma is a well known but very rare neoplasm in humans. To our knowledge, this is the first report of the mucinous variant of adenocarcinoma of the rete testis in a dog.     

A quantitative assessment of the risk of transmission of bovine spongiform encephalopathy by tallow-based calf milk-replacer

A Monte Carlo simulation model was constructed to assess the risk of BSE transmission to calves by calf milk-replacer (CMR). We assumed that any BSE infectivity in the CMR would be associated with the allowable levels of impurities in tallow used to manufacture the milk-replacer. Simulations used three different levels of impurities, six different distributions of the BSE infectivity titers of CNS tissues and with and without inclusion of specified risk material (SRM). Our results suggest that tallow-based CMR could have been responsible for some BSE infections in nearly all simulations. The reduction in the allowable impurities in tallow and the exclusion of SRM have greatly reduced--but have not eliminated--the risk of BSE transmission by CMR The results of the simulations are associated with much uncertainty.     

Validation of the FAMACHA eye color chart for detecting clinical anemia in sheep and goats on farms in the southern United States

Recent studies on sheep and goat farms in the southern United States indicate that multiple-anthelmintic resistance in Haemonchus contortus is becoming a severe problem. Though many factors are involved in the evolution of resistance, the proportion of the parasite population under drug selection is believed to be the single most important factor influencing how rapidly resistance develops. Therefore, where prevention of resistance is an important parallel goal of worm control, it is recommended to leave a portion of the animals untreated. Recently, a novel system called FAMACHA was developed in South Africa, which enables clinical identification of anemic sheep and goats. When H. contortus is the primary parasitic pathogen, this system can be applied on the farm level to reduce the number of treatments administered, thereby increasing the proportion of the worm population in refugia. Since most studies validating the FAMACHA method have been performed in South Africa, it is important that the method be tested in other regions before its use is broadly recommended. We performed a validation study of FAMACHA by testing the system in sheep (n = 847) and goats (n = 537) of various breeds and ages from 39 farms located in Arkansas, Georgia, Louisiana, Florida, and the US Virgin Islands. The color of the ocular conjunctiva of all animals were scored on a 1-5 scale using the FAMACHA card, and blood samples were collected from each animal for determination of packed cell volume (PCV). Fecal samples were also collected from a majority of the animals tested for performance of fecal egg counts (FEC). Correlations between PCV and eye scores, PCV and FEC, and FEC and eye scores were all highly significant for both sheep and goats (P < 0.001). Data for both FAMACHA scores and PCV were evaluated using two separate criteria for anemia: eye score values of 3, 4 and 5 or 4 and 5, and PCV values of < or =19 or < or =15 were considered anemic. Specificity was maximized when eye score values of 4 and 5 were considered anemic and PCV cut off for anemia was < or =19, but sensitivity was low. In contrast, sensitivity was 100% for both sheep and goats when eye score values of 3, 4 and 5 were considered anemic and PCV cut off was < or =15, but specificity was low. In both sheep and goats, predictive value of a negative was greater than 92% for all anemia and eye score categories, and was greater than 99% for both eye score categories when an anemia cutoff of < or =15 was used. Predictive value of a positive test was low under all criteria indicating that many non-anemic animals would be treated using this system. However, compared to conventional dosing practices where all animals are treated, a large proportion of animals would still be left untreated. These data indicate that the FAMACHA method is an extremely useful tool for identifying anemic sheep and goats in the southern US and US Virgin Islands. However, further studies are required to determine optimal strategies for incorporating FAMACHA-based selective treatment protocols into integrated nematode control programs.     

Effect of dexamethasone treatment on the immune response of Gulf Coast Native lambs to Haemonchus contortus infection

Neonatal and weaner Gulf Coast Native (Native) lambs were studied to determine whether an immunological basis underlies their natural resistance to Haemonchus contortus infection. Neonatal Native lambs (n = 8) and weaner Native lambs (n = 15) were randomly assigned to a treatment or a control group. Lambs in the treatment group received dexamethasone by intramuscular injection three times a week for 10 weeks (neonatal) and 15 weeks (weaners). All lambs were monitored for fecal egg count (FEC), blood packed cell volume (PCV), and white blood cell differential counts on a weekly basis for the duration of the studies. Neonatal lambs were kept on pasture with their dams and weaner lambs were dewormed at weaning and kept in pens where they received trickle infections. Serum antibody titers to H. contortus whole worm antigen (WWA) were determined using ELISA. Lymphocyte proliferation assays on peripheral blood mononuclear cells (PBMC) were done to assess lymphocyte function. All lambs were vaccinated with killed Brucella abortus strain 19 to assess the effect of dexamethasone treatment on antibody response. All lambs were necropsied at the end of each study to recover the contents of the gastrointestinal tract for nematode enumeration and identification. The results showed that mean FEC and mean PCV of the treatment group was significantly higher and lower, respectively, than in the control group in both neonatal and weaner lambs from weeks 6 and 5, respectively. At necropsy, total nematode count was significantly higher in treatment groups than in the control groups. Serum antibody titers to H. contortus WWA were significantly lower in treated groups than in control groups. Treatment groups showed a consistent depression in lymphocyte percentage being significantly lower from week 6 in both neonatal and weaner lambs. No differences were found in the response of PBMC to mitogen stimulation between the groups. Lambs in the control groups showed strong positive brucellosis card tests and the treatment groups did not. Dexamethasone treatment resulted in depression of the immune response and loss of natural resistance of Native lambs to H. contortus infection. The results of these studies suggest that some aspects of the immune response may underlie the natural resistance of Native sheep to H. contortus infection.     

The use of KIT and tryptase expression patterns as prognostic tools for canine cutaneous mast cell tumors

Cutaneous mast cell tumors (MCTs) are one of the most common tumors in dogs. Currently, prognostic and therapeutic determinations for MCTs are primarily based on the histologic grade of the tumor, but a vast majority of MCTs are of an intermediate grade, and the prognostic relevance is highly questioned. A more detailed prognostic evaluation, especially of grade 2 canine MCTs, is greatly needed. To evaluate the prognostic significance of KIT and tryptase expression patterns in canine cutaneous MCTs, we studied 100 cutaneous MCTs from 100 dogs that had been treated with surgery only. The total survival and disease-free survival time and the time to local or distant recurrence of MCTs were recorded for all dogs. Using immunohistochemistry, 98 of these MCTs were stained with anti-KIT and antitryptase antibodies. Three KIT- and three tryptase-staining patterns were identified. The KIT-staining patterns were identified as 1) membrane-associated staining, 2) focal to stippled cytoplasmic staining with decreased membrane-associated staining, and 3) diffuse cytoplasmic staining. The tryptase-staining patterns were identified as 1) diffuse cytoplasmic staining, 2) stippled cytoplasmic staining, and 3) little to no cytoplasmic staining. Based on univariate and multivariate survival analysis, increased cytoplasmic KIT staining was significantly associated with an increased rate of local recurrence and a decreased survival rate. The tryptase-staining patterns were not significantly associated with any survival parameter. On the basis of these results, we propose a new prognostic classification of canine cutaneous MCTs, according to their KIT-staining pattern, that can be used for the routine prognostic evaluation of canine cutaneous MCTs.     

Pharmacokinetics of enrofloxacin in neonatal kittens

OBJECTIVE: To determine the pharmacokinetics of enrofloxacin in neonatal kittens and compare the pharmacokinetics of enrofloxacin in young and adult cats. ANIMALS: 7 adult cats and 111 kittens (2 to 8 weeks old). PROCEDURE: A single dose of 5 mg of enrofloxacin/kg was administered to adults (i.v.) and kittens (i.v., s.c., or p.o.). Plasma concentrations of enrofloxacin and its active metabolite, ciprofloxacin, were determined. RESULTS: The half-life of enrofloxacin administered i.v. in 2-, 6-, and 8-week-old kittens was significantly shorter and its elimination rate significantly greater than that detected in adults. The apparent volumes of distribution were lower at 2 to 4 weeks and greater at 6 to 8 weeks. This resulted in lower peak plasma concentration (Cmax) at 6 to 8 weeks; however, initial plasma concentration was within the therapeutic range after i.v. administration at all ages. Compared with i.v. administration, s.c. injection of enrofloxacin in 2-week-old kittens resulted in similar Cmax, half-life, clearance, and area under the curve values. Enrofloxacin administered via s.c. injection was well absorbed in 6- and 8-week-old kittens, but greater clearance and apparent volume of distribution resulted in lower plasma concentrations. Oral administration of enrofloxacin resulted in poor bioavailability. CONCLUSIONS AND CLINICAL RELEVANCE: In neonatal kittens, i.v. and s.c. administration of enrofloxacin provided an effective route of administration. Oral administration of enrofloxacin in kittens did not result in therapeutic drug concentrations. Doses may need to be increased to achieve therapeutic drug concentrations in 6- to 8-week-old kittens.