Sub-cellular localisation of alkaline phosphatase activity in the cytoplasm of tammar wallaby (Macropus eugenii) neutrophils and eosinophils

Alkaline phosphatase (ALP) has been used in studies of neutrophil morphology and function as a marker for identifying different granule populations. In human neutrophils, ALP is found within secretory vesicles, a rapidly mobilisable vesicle population important for upregulating membrane receptors during early activation. Intra-cellular ALP activity in the heterophils of rabbits and guinea pigs, in contrast, is found only in secondary granules. The neutrophils and eosinophils of tammar wallabies (Macropus eugenii) have previously been reported to contain large amounts of ALP activity when stained using routine cytochemical techniques. To define the subcellular location of ALP in this species, cell suspensions were examined using cerium chloride cytochemistry and transmission electron microscopy (TEM). ALP was found in 2 distinct cytoplasmic compartments. One compartment displayed morphology consistent with a subpopulation of secondary granules while a second tubulo-vesicular population appeared similar to the secretory vesicles of human neutrophils. Thin tubular vesicles containing ALP were also identified within the cytoplasm of tammar wallaby eosinophils. Large numbers of ALP-containing vesicles have not been recognised previously in eosinophils and this may represent a novel cytoplasmic compartment. In both cell types, ALP-containing structures showed alteration in morphology following stimulation with N-formyl-Met-Leu-Phe (fMLP) and PMA.     

Effect of anticonvulsant dosages of potassium bromide on thyroid function and morphology in dogs

A placebo-controlled experiment was performed to evaluate the effect of potassium bromide on the canine thyroid gland. Basal total thyroxine, free thyroxine, and basal thyrotropin serum concentrations were evaluated over a 6-month period in potassium bromide-treated and control dogs. A thyrotropin-releasing hormone stimulation test was also performed in all dogs at the beginning and conclusion of the study. Thyroid histopathology was compared between treated and control dogs at the end of the study. No difference was detected in any parameter between the two groups at the end of the study. A decline in thyroid hormone concentrations over the course of the study did occur in both groups of dogs. Potassium bromide does not appear to have a significant effect on canine thyroid function or morphology.     

Risk factors associated with development of seizures after use of iohexol for myelography in dogs: 182 cases (1998)

OBJECTIVE: To determine prevalence of seizures after use of iohexol for myelography and identify associated risk factors in dogs. DESIGN: Retrospective study. ANIMALS: 182 dogs that received iohexol for myelography in 1998. PROCEDURE: Medical records were reviewed for age, breed, sex, weight, dose and total volume of iohexol, injection site, number of injections, lesion type and location, total duration of anesthesia, duration from time of iohexol injection to recovery, presence and number of seizures, and whether surgery followed the myelogram. RESULTS: 39 (21.4%) dogs had at least 1 generalized seizure during or after myelography. Injection site was strongly associated with prevalence of seizures, and risk of seizure was significantly higher after cerebellomedullary injections, compared with lumbar injections. Mean total volume of iohexol administered to dogs that had seizures was significantly higher, compared with that administered to dogs that did not have seizures, although dosage did not differ between groups. Weight was significantly correlated with risk of seizure, and dogs that weighed > 20 kg (44 lb) had higher prevalence of seizures than dogs that weighed < 20 kg. CONCLUSIONS AND CLINICAL RELEVANCE: It is preferential to administer iohexol via the L5-6 intervertebral space to minimize the risk of seizures. Higher prevalence of seizures in large dogs, compared with smaller dogs, may be caused by administration of larger total volumes of contrast agent per volume of CSF.     

Presumptive pseudohypoaldosteronism secondary to chronic urinary tract obstruction from sloughed urinary bladder mucosa and urinary tract infection in a cat

Objective – To describe a case of presumptive secondary pseudohypoaldosteronism (PHA) in a cat with urinary tract infection and chronic urethral obstruction. The obstruction was believed to have resulted from sloughed urinary bladder mucosa secondary to pressure necrosis. Case Summary – A 5‐year‐old, 4 kg, castrated male Siamese cat presented for vomiting and stranguria. Medical history included a perineal urethrostomy for urethral obstruction. Physical examination revealed a large, painful, nonexpressible urinary bladder. Point‐of‐care testing demonstrated electrolyte derangements consistent with a postrenal azotemia and metabolic acidosis. Results of urine culture was positive for bacterial growth. Diagnostic imaging revealed presence of retroperitoneal fluid, marked urinary bladder wall thickening, bilateral hydroureter, mild bilateral pyelectasia, and small nephroliths. The patient was treated for a urinary tract obstruction and infection. In the 3 weeks following initial discharge, the patient was evaluated on multiple occasions for lethargy, intermittent vomiting, inappropriate urination, and progressive polyuria and polydipsia. Although the urinary bladder was easily expressed during repeat examinations, it was persistently distended and subjectively thickened upon palpation. Repeat ultrasound of the urinary tract showed evidence of sloughed tissue in the bladder lumen, likely secondary to chronic urethral obstruction and pressure necrosis. A cystotomy was performed to remove the necrotic tissue, and a revised perineal urethrostomy was done due to a partial urethral stricture. Bladder biopsies were obtained at this time. Postoperatively, the cat was reported by the owners to be urinating normally but continued to be polyuric and polydipsic in the week following discharge. One week after surgery, the cat presented in hypovolemic shock with laboratory findings consistent with a presumptive diagnosis of secondary PHA. New or Unique Information Provided – PHA has not been reported previously in a cat. This case report suggests that aldosterone resistance should be considered in cats with consistent laboratory findings and a history of documented obstructive uropathy and urinary tract infection.     

Detecting and quantifying marijuana metabolites in serum and urine of 19 dogs affected by marijuana toxicity

Veterinarians diagnose marijuana toxicity based on clinical signs and history, or in conjunction with an over-the-counter (OTC) human urine drug screen. With the legalization of recreational marijuana use becoming more prevalent in the United States, a more accurate test to aid in the diagnosis of canine marijuana toxicity is needed. We collected urine and serum samples from 19 dogs with confirmed or suspected marijuana toxicosis from multiple veterinary hospitals and analyzed them with a novel UPLC-MS/MS method. Calibrations from 0.1 to 100 ng/mL and QC materials were prepared. Samples were extracted, purified, and eluted with solid-phase extraction. Urine samples were tested with an OTC human urine drug screen. The limit of detection (LOD) and lower limit of quantification (LLOQ) ranges for marijuana metabolites in serum were 0.05–0.25 ng/mL and 0.1–0.5 ng/mL, respectively. In urine, the LOD and LLOQ ranges for the metabolites were 0.05–0.1 ng/mL and 0.1–0.5 ng/mL, respectively. In serum, median and range of metabolite concentrations (ng/mL) detected included: THC, 65.0 (0.14–160); 11-OH-Δ⁹-THC, 4.78 (1.15–17.8); 11-nor-9-carboxy-Δ⁹-THC, 2.18 (0.71–7.79); CBD, 0.28 (0.11–82.5); and THC-glucuronide, 2.05 (0.72–18.3). In the 19 urine samples, metabolite: creatinine (ng: mg) values detected included: THC, 0.22 (0.05–0.74); 11-OH-Δ⁹-THC, 0; 11-nor-9-carboxy-Δ⁹-THC, 1.32 (0.16–11.2); CBD, 0.19 (0.12–0.26); THC-COOH-glucuronide, 0.08 (0.04–0.11); and THC-glucuronide, 0.98 (0.25–10.7). Twenty of 21 urine samples tested negative for THC on the urine drug screen. All 19 serum samples contained quantifiable concentrations of THC using our novel UPLC-MS/MS method. Utilizing a UPLC-MS/MS method can be a useful aid in the diagnosis of marijuana toxicosis in dogs, whereas using an OTC human urine drug test is not a useful test for confirming marijuana exposure in dogs because of the low concentration of THC-COOH in urine.     

Recombinant Iss as a potential vaccine for avian colibacillosis

Avian pathogenic Escherichia coli (APEC) cause colibacillosis, a disease which is responsible for significant losses in poultry. Control of colibacillosis is problematic due to the restricted availability of relevant antimicrobial agents and to the frequent failure of vaccines to protect against the diverse range of APEC serogroups causing disease in birds. Previously, we reported that the increased serum survival gene (iss) is strongly associated with APEC strains, but not with fecal commensal E. coli in birds, making iss and the outer membrane protein it encodes (Iss) candidate targets for colibacillosis control procedures. Preliminary studies in birds showed that their immunization with Iss fusion proteins protected against challenge with two of the more-commonly occurring APEC serogroups (O2 and O78). Here, the potential of an Iss-based vaccine was further examined by assessing its effectiveness against an additional and widely occurring APEC serogroup (O1) and its ability to evoke both a serum and mucosal antibody response in immunized birds. In addition, tissues of selected birds were subjected to histopathologic examination in an effort to better characterize the protective response afforded by immunization with this vaccine. Iss fusion proteins were administered intramuscularly to four groups of 2-wk-old broiler chickens. At 2 wk postimmunization, chickens were challenged with APEC strains of the O1, O2, or O78 serogroups. One week after challenge, chickens were euthanatized, necropsied, any lesions consistent with colibacillosis were scored, and tissues from these birds were taken aseptically. Sera were collected pre-immunization, postimmunization, and post-challenge, and antibody titers to Iss were determined by enzyme-linked immunosorbent assay (ELISA). Also, air sac washings were collected to determine the mucosal antibody response to Iss by ELISA. During the observation period following challenge, 3/12 nonimmunized chickens, 1/12 chickens immunized with 10 microg of GST-Iss, and 1/12 chickens immunized with 50 microg of GST-Iss died when challenged with the O78 strain. No other deaths occurred. Immunized chickens produced a serum and mucosal antibody response to Iss and had significantly lower lesion scores than nonimmunized chickens following challenge, regardless of the challenge strain. This study expands on our previous report of the value of Iss as an immunoprotective antigen and demonstrates that immunization with Iss can provide significant protection of chickens against challenge with three different E. coli strains.     

Airway inflammation is associated with mucous cell metaplasia and increased intraepithelial stored mucosubstances in horses

This study was performed to determine if a peripheral sample of lung from the site where biopsy is conducted is representative of the rest of the lung and to investigate the relationship between airway inflammation and intraepithelial mucous production in the peripheral airways. Lung parenchyma samples were collected from five different regions of the lung in five control and five heaves-affected horses. Horse groups were defined by clinical response to stabling. Tissue sections were used for semi-quantitative scoring of lesions, to count the number of airways, to quantify the amount of stored mucosubstances (Vs) within the epithelium, and to count the number of epithelial cells in terminal airways. No significant differences were found between lung regions or between groups of horses. Lack of regional differences in airway structures means that a biopsy sample can be used for diagnosis and investigation of diffusely distributed diseases. Airway inflammation was correlated with mucous cell metaplasia and Vs. Therefore, in horses, mucus accumulation is partly caused by increased number of mucous cells and is associated with airway inflammation. Therapy targeted to reduce airway inflammation will help reduce the excessive mucous accumulation in horses.     

Characteristics of free-roaming cats and their caretakers

OBJECTIVES: To describe the characteristics of unowned, free-roaming cats and their caretakers who participated in a trap-neuter-return (TNR) program and to determine the effect of the program on free-roaming cat colonies. DESIGN: Prospective study. SAMPLE POPULATION: 101 caretakers of 920 unowned, free-roaming cats in 132 colonies in north central Florida. RESULTS: Most (85/101; 84%) caretakers were female. The median age was 45 years (range, 19 to 74 years). Most (89/101; 88%) caretakers owned pets and of those, most (67/101; 66%) owned cats. The major reasons for feeding free-roaming cats were sympathy and love of animals. Most caretakers reported that the cats they cared for were too wild to be adopted, but many also reported that they considered the cats to be like pets. The total surveyed cat population was 920 before participation in TNR and 678 after TNR. Mean colony size was 7 cats before TNR and 5.1 cats after TNR. Most cats lived on the caretaker's property. At the time of the survey, 70% (644/920) of the cats had been neutered. CONCLUSIONS AND CLINICAL RELEVANCE: The decrease in the surveyed free-roaming cat population was attributed to a reduction in births of new kittens, adoptions, deaths, and disappearances. Recognition of the human-animal bond that exists between caretakers and the feral cats they feed may facilitate the development of effective control programs for feral cat populations.