Clonality in Sclerotinia sclerotiorum on Infected Cabbage in Eastern North Carolina

ABSTRACT Eighty-four isolates of Sclerotinia sclerotiorum from four cabbage production fields in North Carolina and 16 isolates from an experimental cabbage field plot in Louisiana were DNA-fingerprinted and tested for mycelial compatibility. In a comparison with 594 unique DNA fingerprints of S. sclerotiorum from Canadian canola, no fingerprints were shared among Canadian, North Carolina, and Louisiana populations. DNA fingerprints from the North Carolina sample were distinctive from those of the Canadian and Louisiana samples, with significantly more hybridizing fragments in the 7.7- to 18-kilobase range. Forty-one mycelial compatibility groups (MCGs) and 50 unique DNA fingerprints were identified from the North Carolina sample. Three MCGs and three fingerprints were identified from the Louisiana sample. From the North Carolina sample, 32 MCGs were each associated with a unique fingerprint; of these, there were 11 clones (i.e., cases in which two or more isolates belonged to the same MCG and shared the same DNA fingerprint). Six clones sampled from two or more fields represented approximately 29% of the total sample (24 of 84 isolates), with six clones recovered from fields 75 km apart. There were 10 cases in which one MCG was associated with more than one DNA fingerprint and two cases in which one DNA fingerprint was associated with more than one MCG. The small sample from Louisiana was strictly clonal. The North Carolina sample had a clonal component, but deviated from one-to-one association of MCG with DNA fingerprint to an extent consistent with more recombination or transposition than the other two populations sampled.     

Pyothorax in 26 cats: clinical signs, laboratory results and therapy (2000-2007)

Common historical findings in 26 cats with pyothorax were dyspnea (85%), lethargy (65%), and anorexia (62%), common clinical findings were dyspnea (89%), tachypnea (73%), fever (39%), and hypothermia (27%). Frequent laboratory abnormalities were leukocytosis (68%) with a left shift (100%), anemia (65%), hypoalbuminemia (91%), hyperglobulinemia (86%), hyperbilirubinemia (60%) as well as azotemia (52%). The thoracic effusion was purulent in all cats, the thoracic fluid evaluation (n = 19) met the criteria for an inflammatory exudate (protein 32-63 g/l, median 44; cell count 54.4-390 x 10(9)/l, median 100). Cytological analysis revealed bacteria in 21 of 24 cats. From 16 cats 13 different genera were isolated, 5 cultures were without bacterial growth. In 12.5% of the cats a single population of bacteria was identified, and in 87.5% a mixture of 2-4 different bacterial species were cultured. Obligate anaerobic bacteria were the most common isolates (70%) followed by facultative anaerob (22.5%) and aerob growing (7.5%) bacteria. Bacterial culture most often yielded Fusobacteria spp., Prevotella spp., Pasteurella spp., Porphyromonas spp. and Bacteroides spp. Three cats were euthanized at the day of presentation. A thoracic lavage was performed in 22 cats; mechanical complications with the chest tubes occured in 4 cats. Thoracotomy was performed in 2 cats. Of the 23 treated cats 17 survived (74%).     

Methicillin-resistant Staphylococcus aureus (MRSA) isolated from small and exotic animals at a university hospital during routine microbiological examinations

Clinical specimens of small animals (n=869) were screened for the occurrence of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA; MRSA) during routine microbiological examinations, and results were confirmed by a multiplex PCR strategy. The genetic relatedness of all mecA-positive S. aureus isolates was further investigated by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), PCR for Panton-Valentine leukocidine genes (PVL) and staphylococcal cassette chromosome mec-typing (SCCmec). A total of 61 S. aureus isolates were found during a 20-month period of investigation, 27 (44.3%) of them harbouring the mecA gene for methicillin-resistance. The majority of MRSA were isolated in specimens from dogs (n=18) and cats (n=4). One guinea pig and one rabbit were found to be positive for an MRSA infected site. Similarly, three exotic animals, a turtle, a bat and a parrot, were found to be infected with MRSA. PFGE and MLST analysis revealed a certain genotype ("A" and "A-1") dominating the isolate collection (23 of 27). Furthermore, one isolate showed homologous PFGE pattern to the German epidemic strain Barnim ("BE") and another one ("BE-1") was considered to be closely related. A third genotype ("B") was detected in two cases. Two different sequence types (ST) were identified among the 27 MRSA isolates. PFGE type "A" and both strains related to the Barnim epidemic strain were assigned to ST22, whereas ST239 was associated to PFGE profile "B". The present data show that certain MRSA genotypes are capable of infecting a wide spectrum of small and exotic animals, especially in clinical facilities.     

Primary immune-mediated thrombocytopenia in 30 dogs (1997-2003)

During a 6-year period, primary (idiopathic) immune-mediated thrombocytopenia was retrospectively evaluated in 30 dogs. Ages of the dogs ranged from 3 months to 10 years (median 4 years); female dogs were markedly overrepresented with 73%. Clinical examination revealed hemorrhages in 70% of the dogs. Platelet counts ranged from 0 to 111,000/microL (median 8000/microL); 77% of the dogs had platelet counts <30,000/microL. Seventeen dogs were anemic (hematocrit 9% to 36%; median 31%). Immunosuppressive therapy was performed in all but one dog. The recurrence rate of 19 dogs that were followed over an extended period (112 to 1684 days; median 340 days) was 26%. Twenty-nine (97%) dogs survived 14 days after initial presentation, and 27 (93%) dogs survived at least the following 15 to 1684 days (median 220 days).     

Rapid calcitonin response to experimental hypercalcemia in healthy horses

Calcium has important physiological functions, and disorders of calcium homeostasis are frequent in horses. We have made important progress understanding equine calcium homeostasis; however, limited information on equine calcitonin (CT) is available, in part because of the lack of validated CT assays. To determine the CT response to high ionized calcium (Ca²⁺) concentrations in healthy horses, we induced hypercalcemia in 10 healthy horses using a calcium gluconate 23% solution (5 mg/kg; 120 mL/500 kg horse) infused over 4 min. Four horses were infused with 120 mL of 0.9% NaCl and used as controls. We validated a human-specific CT radioimmunoassay for use in horses. Serum Ca²⁺ concentrations increased from 6.2 ± 0.3 mg/dL to 9.9 ± 0.5 mg/dL (4 min; P < 0.01). Serum CT increased from 16.7 ± 8.0 pg/mL to 87.1 ± 55.8 pg/mL at 2 min, and 102.5 ± 51.1 pg/mL at 4 min (P < 0.01). Serum CT returned to baseline by 20 min, whereas serum Ca²⁺ returned to baseline by 40 min. Of interest, CT concentrations returned to baseline despite hypercalcemia, suggesting thyroid gland C-cell CT depletion. Resting CT values higher than 40 pg/mL were considered abnormally elevated. No significant changes in serum Ca²⁺ or CT concentrations were found in control horses. The coefficients of variation for the CT radioimmunoassay were lower than 11.9%. We conclude that the equine thyroid gland C-cell responds quickly to changes in extracellular Ca²⁺ concentrations by secreting large quantities of CT into the systemic circulation, indicating that CT is important in equine calcium homeostasis. The human CT radioimmunoassay can be used to measure changes in equine CT.     

Primary immune-mediated hemolytic anemia in 19 cats: diagnosis, therapy, and outcome (1998-2004)

Immune-mediated hemolytic anemia (IMHA) occurs less frequently in cats than in dogs. The value of the Coombs' test (CT) has been questioned, but detailed surveys of its use are lacking. The objective of this study was to describe 19 cats with primary IMHA (pIMHA) and to examine the diagnostic value of the direct CT. The CT was performed in 92 cats; it was negative in 5 healthy, in 9 sick nonanemic, and in 55 cats with different types of anemia. The CT was positive in 18 anemic cats (2 feline leukemia virus (FeLV) positive, 1 with cholangiohepatitis, 15 with no underlying disease). Moreover, agglutination persisted after saline washing in 5 anemic cats (1 lymphoma, 4 pIMHA). Inclusion criteria for pIMHA were a positive CT (15) or persistent agglutination (4), and the exclusion of other diseases. The age of the 19 cats ranged from 0.5 to 9 years (median, 2 years); male cats were overrepresented. The PCV on admission was 6-22% (median, 12%). The anemia was nonregenerative in 11 cats. Additional abnormal laboratory results were leukocytosis (2), lymphocytosis (6), hyperbilirubinemia (13), hyperglobulimemia (10), and increased liver enzyme activities (10). Initial treatment consisted of blood transfusions (10), crystalloids (11), prednisolone (19), antibiotics (19), and H2-blockers (11). Four of 17 cats were euthanized 9, 63, 240 and 2,160 days after initial presentation (mortality rate, 23.5%). Relapses were reported in 5 of 16 cases (31%). Thus, pIMHA appears to occur more frequently than recognized previously, with a more favorable prognosis in cats than in dogs. The CT was useful in identifying immune-mediated pathogenesis.