Investigations into the growth and formation of biofilm by at temperatures encountered during infection.

The genus comprises unique atypical spirochete bacteria that includes the etiological agent of leptospirosis, a globally important zoonosis. Biofilms are microecosystems composed of microorganisms embedded in a self-produced matrix that offers protection against hostile factors. Leptospires form biofilms in rice fields and unsanitary urban areas, and while colonizing rodent kidneys.

Circulating Foamy Macrophages in the Golden Syrian Hamster (Mesocricetus auratus) Model of Leptospirosis.

Leptospirosis is a world-wide zoonotic disease caused by pathogenic Leptospira and can be asymptomatic or can cause clinical signs ranging from influenza-like to multi-organ failure and death in severe cases. While species and strain specificity can play a major role in disease presentation, the hamster is susceptible to most leptospiral infections and is the model of choice for vaccine efficacy testing. During evaluation of blood smears from hamsters challenged with different species and strains of Leptospira, a circulating population of large, mononuclear, lipid-filled cells, most similar to foamy macrophages (FMs), was detected.

Disseminated aspergillosis in a dog due to Aspergillus alabamensis.

Disseminated aspergillosis is uncommon in dogs and often associated with Aspergillus terreus. A case of disseminated disease in an English springer spaniel is reported from which Aspergillus alabamensis was recovered by culture and identified by molecular means suggesting a potential role for this agent as a primary pathogen of dogs.

Heterophil functional responses to dietary immunomodulators vary in genetically distinct chicken lines.

The effect of dietary supplementation of immunomodulators on in vitro chicken heterophil function was investigated using three diverse genetic lines of chickens (broiler, Fayoumi, and Leghorn). Dietary supplementation with β-glucan, ascorbic acid, and corticosterone was fed from 8 to 11 weeks of age. Heterophil function was evaluated weekly during supplementation using phagocytosis, bacterial killing, and heterophil extracellular traps (HETs)-DNA release.

Genetic control of chicken heterophil function in advanced intercross lines: associations with novel and with known Salmonella resistance loci and a likely mechanism for cell death in extracellular trap production.

Heterophils, the avian polymorphonuclear leukocyte and the counterpart of mammalian neutrophils, generate the primary innate response to pathogens in chickens. Heterophil performance against pathogens is associated with host disease resistance, and heterophil gene expression and function are under genetic control. To characterize the genomic basis of heterophil function, heterophils from F(13) advanced intercross chicken lines (broiler × Leghorn and broiler × Fayoumi) were assayed for phagocytosis and killing of Salmonella enteritidis, oxidative burst, and extracellular trap production.

Protecting animal and human health and the nation's food supply through veterinary diagnostic laboratory testing.

The current detection system for animal diseases requires coordination between veterinarians; veterinary medical laboratories; and state, federal, and international agencies, as well as associated private sector industries. Veterinary clinical pathologists in clinical and governmental laboratories often have responsibilities and expertise in one or more laboratory disciplines involved in diagnosing zoonotic and/or emerging diseases and diseases exotic to the United States that are important to animal and human health and the nation's food supply. The knowledge and roles of all veterinary laboratory professionals are vital to detect, monitor, and confirm diseases and conditions that affect animal and human health and the nation's animal food supply.

Proportion of circulating chicken heterophils and CXCLi2 expression in response to Salmonella enteritidis are affected by genetic line and immune modulating diet.

Genetic line and diet affect chicken heterophil activity and gene expression, and the combination of these factors can enhance disease resistance. This study evaluated the effects of immune modulating diets on heterophil/lymphocyte (H/L) ratio and heterophil chemokine expression in distinct genetic lines. Fayoumi and Leghorn chickens were fed a basal diet or immune modulating diets enhanced with β-glucans, ascorbic acid, or corticosterone.

Use of rapid cytochemical staining to characterize fish blood granulocytes in species of special concern and determine potential for function testing.

Studies of innate immunity in fish species of special concern are essential for better understanding of their health status during hatchery rearing conditions. The cytochemical and morphological characterizations of blood granulocytes have been used to provide information about phylogenetic differences and determine the potential use of neutrophil function assays. Rapid, simple, cytochemical staining kits used routinely for staining mammalian granulocytes have been used to characterize granulocytes from blood of four fish species: Arctic grayling, cutthroat trout, June sucker, and shovelnose sturgeon.

Chicken heterophils from commercially selected and non-selected genetic lines express cytokines differently after in vitro exposure to Salmonella enteritidis.

Resistance to pathogens such as Salmonella enteritidis (SE) is a heritable trait important in maintaining the health of chickens and reducing bacterial contamination of poultry products. In chickens, heterophils act as the first responders to bacterial infections and are, therefore, responsible for initiating the immune response against SE challenge. This study measured mRNA expression of several immune response genes [interleukin-6 (IL-6), IL-10, transforming growth factor-beta4 (TGF-beta4), granulocyte macrophage-colony stimulating factor (GM-CSF), and Toll-like receptor-4 (TLR-4)] by heterophils from broiler, Leghorn, and Fayoumi chickens, either non-stimulated or stimulated in vitro with SE using quantitative reverse-transcriptase PCR.

Chicken heterophil extracellular traps (HETs): novel defense mechanism of chicken heterophils.

Recent findings in mammals and fish have revealed that neutrophil nuclear material associated with cytoplasmic granular content is released in the form of neutrophil extracellular traps (NETs) that can trap and kill invading microorganisms in vitro and in vivo. To determine if a similar mechanism is present in chicken heterophils, hydrogen peroxide (H(2)O(2)) and phorbol myristate acetate (PMA) were used for stimulation of blood-derived heterophils. Stimulated heterophils produced structures that were characterized using immunocytochemistry and confocal microscopy as heterophil extracellular traps (HETs).

Evaluation of the use of subcutaneous implantable vascular access ports in feline blood donors.

Objective: To compare the ease and effects of collecting blood from cats by use of subcutaneous totally implantable vascular access ports (VAPs) with collection via conventional jugular phlebotomy.

Design: Prospective randomized experimental study.

Animals: 8 healthy cats.

Fish cast NETs: neutrophil extracellular traps are released from fish neutrophils.

Neutrophil extracellular traps (NETs), which are extracellular DNA structures released from neutrophils, are described and characterized for the first time in fish using fluorescent confocal microscopy. Confocal images of fish neutrophil suspensions stained with 6'-diamino-2-phenylindole, dihydrochloride DNA fluorescent stain (DAPI) revealed the presence of NETs which appeared as fibrous structures connecting several cells. Co-localization of NETs with neutrophil granular proteins and actin was investigated using specific antibodies and probes.

Zebrafish (Danio rerio) whole kidney assays to measure neutrophil extracellular trap release and degranulation of primary granules.

The zebrafish (Danio rerio) is an excellent model system for studies in developmental biology, genetics, and toxicology, and is increasingly gaining importance in the field of immunology. The use of whole zebrafish kidneys as source of neutrophils for degranulation assays and detection of neutrophil extracellular traps is described for the first time. Neutrophils from zebrafish kidneys released neutrophil extracellular traps (NETs) and myeloperoxidase (MPO) upon stimulation with calcium ionophore, phorbol myristate acetate, and beta-glucan.

Immunomodulatory effects of beta-glucan on neutrophil function in fathead minnows (Pimephales promelas Rafinesque, 1820).

Stimulatory effects of yeast beta-1,3-1,6-glucans on neutrophils have long been recognized, but effects of glucans on degranulation of primary granules in fish neutrophils have not been previously reported. Neutrophil function was monitored during in vitro and in vivo application of glucans to non- (NS), acute- (AS) and chronically stressed (CS) fish. beta-Glucan proved to be a strong and quick (80%, 2 min) stimulant of degranulation.

Experimental canine leptospirosis caused by Leptospira interrogans serovars pomona and bratislava.

Objective: To evaluate gross, histopathologic, and serum biochemical findings caused by Leptospira interrogans serovars pomona and bratislava inoculated in dogs.

Animals: Twenty-seven 8-week-old female Beagles.

Procedure: Dogs were randomly assigned to challenge or control groups.

A rapid, direct assay to measure degranulation of primary granules in neutrophils from kidney of fathead minnow (Pimephales promelas Rafinesque, 1820).

A direct, rapid, quantitative colorimetric assay to determine neutrophil primary granule degranulation was adapted for use with fathead minnow kidney neutrophils. The assay measures the exocytosis of myeloperoxidase (MPO) using 3,3',5,5'-tetramethylbenzidine as a substrate. The assay was validated by comparing the total myeloperoxidase content of neutrophil populations obtained from adult cattle, as a known positive, and fish; evaluating the effects of calcium ionophore (CaI), phorbol myristate acetate (PMA), aqueous solution of beta-glucan (MGAQ) and zymosan (Z) with and without cytochalasin B (cyto B) as stimulants of degranulation; determining the kinetics of primary granule exocytosis and detecting changes in degranulation when fish were exposed to stress and anaesthesia with MS-222.

Clinical and pathologic comparison of acute leptospirosis in dogs caused by two strains of Leptospira kirschneri serovar grippotyphosa.

Objective: To develop a method for inducing acute leptospirosis in dogs.

Animals: 31 nine-week-old female Beagles.

Procedure: Beagles were randomly assigned to 2 inoculation groups or a control group.

Keratinizing ameloblastoma in a 9-month-old llama (Lama glama).

A 9-month-old male llama (Lama glama) was presented because of a rapidly growing mass on the right side of the face. Radiographs revealed a marked expansion of the right caudal face region with bone lysis involving the maxilla and the nasal, lacrimal, zygomatic, and palatine bones. Cytologically, the mass consisted of atypical round to polygonal cells with round nuclei and basophilic cytoplasms that formed acini and rows.

The utility of plasma D-dimer to identify thromboembolic disease in dogs.

This prospective study was designed to investigate D-dimer concentrations in clinically healthy dogs, clinically ill dogs without thromboembolic disease (TE), and dogs with TE. The goals of this study were to determine whether the coagulation cascade is activated in nonembolic metabolic and inflammatory conditions and whether differentiation from TE is possible. Group 1 consisted of 30 clinically healthy dogs presented for routine care.

Bronchoalveolar lavage.

Bronchoalveolar lavage is a valuable diagnostic technique when appropriate patient selection is made, which includes diseases that involve the pulmonary parenchyma and patients that can safely undergo general anesthesia. Bronchoalveolar lavage cytology has the potential to provide a definitive diagnosis or additional diagnostic information with a less invasive procedure than lung aspiration or thoracotomy. Limitations of accuracy in cell counts and negative findings are considerations when interpreting bronchoalveolar lavage fluid results.

Analysis of transtracheal aspirates and pleural fluid from clinically healthy llamas (Llama glama).

Seventeen clinically healthy adult llamas were used to study the characteristics of transtracheal aspirates (TTA) and pleural fluid samples. Results of complete blood counts, fibrinogen determination and thoracic radiographs were within normal limits prior to sampling. Cytologic evaluation of TTA revealed the majority of cells were vacuolated macrophages (60-100%), with 0-40% neutrophils, and fewer lymphocytes (0-1%), eosinophils (0-3%), and ciliated respiratory epithelial cells (0-10%).