Serovar identification, antimicrobial sensitivity, and virulence of Avibacterium paragallinarum isolated from chickens in Thailand

Avibacterium paragallinarum causes infectious coryza in chickens, an acute respiratory disease that has worldwide economic significance. The objectives of this study were to determine the serovars, antimicrobial resistance, and pathogenicity of A. paragallinarum isolated from chickens in Thailand. Eighteen field isolates of A. paragallinarum were confirmed by PCR. When examined by serotyping in a hemagglutination inhibition test, 10 isolates were serovar A, five isolates were serovar B, and three isolates were serovar C. The susceptibility of the isolates to 16 antimicrobial agents was tested by a disk diffusion method. All isolates were susceptible to amoxicillin-clavulanic acid. There was a high level of resistance to lincomycin and erythromycin. All isolates were resistant to cloxacillin and neomycin. A study of bacterial entry into, and survival within, chicken macrophages showed variation between isolates but no clear connection to serovar. A virulence test was performed by challenging 4-wk-old layers via the nasal route with 400 dl of bacteria (10(8) colony-forming units/ml). Clinical signs were observed daily for 7 days, and the birds were subjected to a postmortem necropsy at 7 days postchallenge. All 18 field isolates caused the typical clinical signs of infectious coryza and could be re-isolated at 7 days after challenge. There was no significant difference in the clinical scores of the isolates except that two isolates (112179 and 102984, serovars A and B, respectively) gave a significantly higher score than did isolate CMU1009 (a serovar A isolate). No correlation between serovar and severity of clinical signs was found.     

Morphology and Neurochemical Expression of Neurons Immunoreactive for the Calcitonin Gene-Related Peptide (CGRP-IR) in the Lamb Ileum

Veterinary Research Communications -     

Cardiorespiratory Effects of Combined Midazolam and Butorphanol in Isoflurane-anesthetized Cats

The cardiorespiratory effects of midazolam (0.1 mg/kg) and butorphanol (0.4 mg/kg) were evaluated in seven cats. Cats were induced in a chamber, removed and intubated, and maintained with isoflurane in 100% oxygen. Cardiorespiratory data were recorded before and 3, 10,20,30,40,50, and 60 minutes after intravenous injection of midazolam-butorphanol. Heart rate, mean arterial pressure, and respiration rate were significantly (p < .05) decreased below baseline at various times throughout the study. Tidal volume and minute ventilation were unchanged. End-tidal carbon dioxide was significantly (p < .05) increased above baseline for 30 minutes. Midazolam-butorphanol administered intravenously induced significant alterations in several cardiorespiratory parameters in isoflurane-anesthetized cats.     

Fatal toxoplasmosis and concurrent Calodium hepaticum infection in Korean squirrels (Tanias sibericus)

Four Korean squirrels (Tanias siberius) imported in Spain from People's Republic of China died 2 days after their arrival at a pet shop. They had neurological signs associated with generalized toxoplasmosis involving brain, lungs, liver, and the heart. Toxoplasma gondii-like tachyzoites and tissue cysts were found in organs of all four squirrels. The protozoa stained positively with T. gondii polyclonal antibodies and were ultrastructurally similar to T. gondii. Calodium (Capillaria) hepaticum infection was found in the liver of one squirrel.     

Disseminate Cryptococcosis in a guenon (Cercopithecus ascanius)

A 23-year-old captive-bred red-tailed guenon (Cercopithecus ascanius) with a brief history of inappetence, lethargy, and seizures was submitted for necropsy. On postmortem examination, multiple cryptococcomas were identified in brain and heart. Cryptococcus neoformans organisms were also identified microscopically in kidney, eye, and pancreas. Fungal yeast formed rare pseudohyphae. The histologic diagnosis of cryptococcosis was confirmed by a positive test for C. neoformans antigen in a serum sample. Immunohistochemical staining confirmed that macrophages were the principal inflammatory cell in brain lesions and often contained phagocytosed yeast. As disseminate cryptococcosis is often associated with immune suppression, serology and immunohistochemical staining for simian immunodeficiency virus were performed but showed no evidence of SIV infection.     

The detection of anthelmintic resistance in nematodes of veterinary importance

Before revised World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines on the detection of anthelmintic resistance can be produced, validation of modified and new methods is required in laboratories in different parts of the world. There is a great need for improved methods of detection of anthelmintic resistance particularly for the detection of macrocyclic lactone resistance and for the detection of resistant nematodes in cattle. Therefore, revised and new methods are provided here for the detection of anthelmintic resistance in nematodes of ruminants, horses and pigs as a basis for discussion and with the purpose that they are evaluated internationally to establish whether they could in the future be recommended by the WAAVP. The interpretation of the faecal egg count reduction test has been modified and suggestions given on its use with persistent anthelmintics and continuous release devices. An egg hatch test for benzimidazole (BZ) resistance is described. A microagar larval development test for the detection of benzimidazole and levamisole resistance provides third stage larvae for the identification of resistant worms. The sensitivity of these two tests can be increased by using discriminating doses rather than LD(50) values. Details are given of a PCR based test for the analysis of benzimidazole resistance in strongyles of sheep and goats, horses and cattle. Although promising for ruminant trichostrongyles, quantitative determination of gene frequency using real time PCR requires further development before PCR tests will be used in the field. Apart from faecal egg count reduction tests there are currently no satisfactory tests for macrocylic lactone resistance despite the great importance of this subject. Except for treatment and slaughter trials there are no validated tests for fasciolicide resistance or for the detection of resistance in cestodes.     

Genotyping of Toll-like receptor 4, myeloid differentiation factor 2 and CD-14 in the horse: an investigation into the influence of genetic polymorphisms on the LPS induced TNF-alpha response in equine whole blood

The inter- and intra-species differences in the response to lipopolysaccharides (LPS) are well recognised in mammalian species. It has been hypothesized that these differences can be attributed to genetic polymorphisms in the components involved in LPS signal transduction. These components include the cluster of differentiation factor 14 (CD-14), a membrane bound protein on the surface of mononuclear cells that recognises LPS and a receptor complex consisting of Toll-like receptor-4 (TLR-4) and myeloid differentiation factor-2 (MD-2). Sequencing of these three proteins in humans and mice revealed that all three are susceptible to polymorphic alterations, influencing the response to LPS. Previous experiments in the horse showed large inter-individual variations in the response to LPS. With the aim to assess this inter-individual variation, we performed a whole blood assay in 10 healthy horses as a functional assay to study the responsiveness to LPS. In 3 out of the 10 horses, LPS-induced TNF-alpha production was significantly lower compared to the overall mean. Subsequently the entire cDNA sequence encoding for the TLR-4, MD-2 and CD-14 protein was documented for each horse. Although mutations were observed in the sequence of TLR-4, these could not be related to an altered response to LPS in the concentration used in this study, as determined in the whole blood assay. Despite the various mutations found in the TLR-4 receptor protein, no alterations could be found in either the MD-2 or CD-14 gene, which are obviously more conserved structures.