Analysis of fenbendazole residues in bovine milk by ELISA

Fenbendazole residues in bovine milk were analyzed by ELISAs using two monoclonal antibodies. One monoclonal antibody (MAb 587) bound the major benzimidazole anthelmintic drugs, including fenbendazole, oxfendazole, and fenbendazole sulfone. The other (MAb 591) was more specific for fenbendazole, with 13% cross-reactivity with the sulfone and no significant binding to the sulfoxide metabolite. The limit of detection of the ELISA method in the milk matrix was 7 ppb for MAb 587 and 3 ppb for MAb 591. Fenbendazole was administered in feed, drench, and paste form to three groups of dairy cattle. Milk was collected immediately before dosing and then every 12 h for 5 days. The ELISA indicated that residue levels varied widely among individual cows in each group. Fenbendazole levels peaked at approximately 12-24 h and declined rapidly thereafter. Metabolites were detected at much higher levels than the parent compound, peaked at approximately 24-36 h, and declined gradually. Residue levels were undetectable by 72 h. The ELISA data correlated well with the total residues determined by chromatographic analysis, but the use of the two separate ELISAs did not afford an advantage over ELISA with the single, broadly reactive MAb 587. The ELISA method could be used to flag high-residue samples in on-site monitoring of fenbendazole in milk and is a potential tool for studying drug pharmacokinetics.     

Location of roosts of the lesser long-eared bat Nyctophilus geoffroyi and Gould's wattled bat Chalinolobus gouldii in a fragmented landscape in south-eastern Australia

The location of roosts of two species of vespertilionid bats, the lesser long-eared bat (Nyctophilus geoffroyi) and Gould’s wattled bat (Chalinolobus gouldii), was investigated in a remnant vegetation-farmland mosaic and adjacent floodplain forest in south-eastern Australia. A total of 45 individuals of N. geoffroyi and 27 C. gouldii were fitted with radio transmitters, which resulted in the location of 139 and 89 roosts respectively. Selection of roosting habitat showed both inter- and intra-specific differences. At the landscape level, locations of roosts used by male and female N. geoffroyi differed markedly. Most males roosted in the farmland mosaic within 3 km (mean 1.9±2.9 km) of where they were captured while foraging in remnant vegetation. In contrast, roost sites of females were predominantly in the floodplain forest, 6-12 km from their capture site in farmland (mean for all females, 6.7±2.9 km). All maternity roosts were in the extensive floodplain forest, 4-10 km from foraging areas. Distances moved by this species were greater than previously recorded and further than predicted by flight morphology. Most C. gouldii roosted in the floodplain forest, 4-10 km from their capture site (6.9±1.6 km). Within the floodplain forest, both species roosted in areas that had higher densities of hollow-bearing trees than generally available; dead hollow-bearing trees for N. geoffroyi and large, live trees for C. gouldii. The scale of movements undertaken by these species means that they can obtain resources from multiple landscape elements. Both species used different habitats for roosting and foraging despite the energetic costs of commuting relatively large distances. Conservation of bat populations in rural environments needs to be considered at the landscape scale, with particular attention to identifying landscape elements that provide key resources.     

A retrospective study of 11 cases of lungworm (Didelphostrongylus hayesi) infection in opossums (Didelphis virginiana).

A juvenile, female North American opossum (Didelphis virginiana) died of verminous pneumonia caused by Didelphostrongylus haysei despite aggressive treatment with oral fenbendazole, corticosteroids, and antibiotics. This prompted a retrospective study of lungworm infection in opossums, during which 19 additional necropsy reports from opossums were reviewed. Including the subject of this report, a total of 11 (55%) of these cases included a diagnosis of lungworm infection. This diagnosis was considered to have contributed to death in eight out of the 11 cases (73%). Histologically, 10 of the 11 (91%) opossums had granulomatous bronchopneumonia with small to moderate numbers of adult nematodes in the airways and parenchyma. Four of the 11 (36%) opossums had free larvae within the parenchyma or terminal airways. Inflammation was usually associated with larvae, degenerating parasites, and nonintact adult nematodes. Superimposed bacterial pneumonia was evident in three animals, and sections of lung examined from all the opossums were characterized by moderate to severe smooth-muscle hyperplasia in airways, including terminal respiratory bronchioles and alveolar ducts. Nine animals had prominent medial smooth-muscle hyperplasia in small- and medium-sized arterioles. Lesions in other organs, particularly in liver, heart, and gastrointestinal tract, were frequently identified. Three animals had concomitant septicemia or bacterial bronchopneumonia (or both), which contributed to the cause of death. Seven animals had gastric nematodosis (Physaloptera sp.), although three of them had been treated with a 14-day course of fenbendazole.     

Blastomycosis in nondomestic felids.

Blastomycosis was diagnosed in six nondomestic felids from eastern Tennessee, including two Asian lions (Panthera leo persicus), one African lion (Panthera leo), one Siberian tiger (Panthera tigris), one cheetah (Acinonyx jubatus), and one snow leopard (Panthera uncia). Clinical signs included lethargy, anorexia, weight loss, dyspnea, sneezing. ataxia, and paresis. Variable nonspecific changes included leukocytosis, monocytosis, moderate left shift of neutrophils, moderate hypercalcemia, hyperproteinemia, and hyperglobulinemia. Thoracic radiographs revealed interstitial and alveolar changes, consolidation or collapse of a lung lobe, bullae formation, and a pulmonary mass. Agar gel immunodiffusion (AGID) serology for Blastomyces dermatitidis was performed in five felids and was positive in three. The tiger had cerebral blastomycosis and was positive for AGID serologic tests of both cerebrospinal fluid and serum. One percutaneous lung aspirate in the snow leopard and one bronchial aspirate in an Asian lion demonstrated B. dermatitidis organisms. whereas tracheal wash samples and a nasal discharge were nondiagnostic in others. Treatment with itraconazole was attempted in four cats. The tiger improved before euthanasia, whereas the others did not survive beyond initial treatments. In four felids, B. dermatitidis was found in the lungs and tracheobronchial lymph nodes associated with a florid pyogranulomatous reaction; the tiger had a pyogranulomatous encephalomyelitis, and the cheetah had a single pulmonary granuloma. Thoracic radiography, cytologic examination of lung lesion aspirates, and B. dermatitidis AGID serology should be performed on clinically ill zoo felids in endemic areas to rule out blastomycosis.     

SONOGRAPHIC EVALUATION OF THE CRANIAL MEDIASTINUM IN SMALL ANIMALS

Clinical records, radiographs, and sonograms of 17 animals presented for possible cranial mediastinal disease were reviewed. Radiographs were evaluated for accuracy in detection of mediastinal masses. Sonography of the mediastinum was performed using a parasternal approach with the animals in sternal or lateral recumbency. The mediastinum was evaluated for the presence of a mass or other disease. Masses were characterized by echogenicity, size, number, and margination. Mediastinal vessels were studied to determine if there was vascular invasion or compression. A comparison of radiographic and sonographic assessment in cranial mediastinal disease indicates sonography adds valuable information in the diagnosis of mediastinal disease. Results of ultrasound-guided, fine-needle aspirate were documented and found to be advantageous in establishing a diagnosis in mediastinal masses.     

Bronchopulmonary Disease in the Cat: Historical, Physical, Radiographic, Clinicopathologic, and Pulmonary Functional Evaluation of 24 Affected and 15 Healthy Cats

The results of clinical and pulmonary functional evaluation of 24 cats with bronchopulmonary disease and 15 healthy cats are presented. Affected cats had historical evidence of excessive reflexes (coughing, sneezing); physical evidence of airway secretions (crackles), obstruction (wheezing), and increased tracheal sensitivity; radiographic evidence of bronchial and interstitial lung disease; and cytological evidence of airway inflammation or mucous secretions. Bacterial isolates from healthy and affected cats were predominantly Gram-negative rods, indicating that bronchi of cats are not always sterile and that normal flora should be considered in interpreting cultures from cats with suspected bronchopulmonary disease. Cats were grouped according to relative disease severity based on scored historical, physical, and radiographic abnormalities. The mean (± standard deviation) baseline lung resistance measurement in healthy cats was 28.9 cm H₂O/L/s (±6.2 cm H₂O/L/s), whereas in mildly, moderately, and severely affected cats it was 38.3 cm H₂O/L/s (±21.5 cm H₂O/L/s), 44.8 cmH₂O/L/s (±7.7 cm H₂O/L/s), and 105.2 cm H₂O/L/s (±66.9 cm H₂O/L/s), respectively. In healthy cats, dynamic lung compliance was 19.8 (±7.4), whereas in mildly, moderately, and severely affected cats it was 14.7 mL/cm H₂O (±3.8 mL/cm H₂O), 17.7 mL/cm H₂O (±6.9 mL/cm H₂O), and 13.0 mL/cm H₂O (±7.9 mL/cm H₂O), respectively. Thus, airway obstruction was present in many of the affected cats. Based on acute response to the bron-chodilator, terbutaline, airway obstruction was partially reversible in many affected cats, although the degree of reversibility varied. Furthermore, based on bronchoprovocation testing, 6 (of 7) affected cats evaluated also had increased airway responsiveness to aerosolized methacholine.     

Potential Corridors for the Rusty Crayfish, <Emphasis Type="Italic">Orconectes Rusticus</Emphasis>, in Northern Wisconsin (USA) Lakes: Lessons for Exotic Invasions

We sampled 35 lakes in northern Wisconsin to determine the presence of Orconectes rusticus, the rusty crayfish, and related this pattern to several parameters pertaining to potential invasion routes that could influence the distribution of these crayfish in the lakes. The presence of rusty crayfish in lakes was positively related to an index of human use and negatively related to the length of stream connections to other lakes containing the crayfish. Humans appear to act as vectors allowing crayfish to travel along discontinuous routes that otherwise would be inaccessible to them, and thus, provide crayfish with spatially discontinuous corridors that increase the probability of movement by channelizing the space between lakes. In contrast, streams correspond closely to the traditional definition of terrestrial corridors, in that they are spatially continuous. This distribution pattern suggests colonization operating via two corridors with two spatial scales, making management of the invasion of rusty crayfish complex.