A Case Report of Subcutaneously Injected Liposomal Cannabidiol Formulation Used as a Compassion Therapy for Pain Management in a Dog.

A 14-year-old intact mixed breed dog (26 kg) was submitted for a novel cannabidiol (CBD) analgesic treatment. The dog was cachectic and had a testicular neoplasia, hip and elbow osteoarthritis and severe cervical pain. Analgesic treatment included canine osteoarthritic supplement, robencoxib and gabapentin. An additional liposomal CBD injectable formulation at 5 mg/kg was administered subcutaneously between the shoulder blades. The dog was monitored using an activity monitoring collar (PetPace), owner wellbeing questionnaire (Canine Brief Pain Inventory; CBPI), pain interactive visual analog scale (iVAS), blood work and CBD plasma concentrations. A week from the injection and up to 3 weeks afterwards the dog had improved CBPI and iVAS pain scores, and increased collar activity scores. CBD was quantified in plasma for 28 days. Due to disease progression, further difficulty to rise and walk, and relapse to pain after 3 weeks, the owners requested a second liposomal CBD injection, which was performed 4 weeks following the first injection using 3 mg/kg dose. Two days later, the dog was found dead in the yard under direct sun, while environmental temperature was 37°C. Major findings on necropsy revealed evidence of heat stroke and severe cervical disc protrusion with spinal hematoma, none related to liposomal CBD. In conclusion, subcutaneous liposomal CBD produced quantifiable CBD plasma concentrations for 28 days and may be an effective additional treatment as part of multimodal pain management in dogs.

Serological cross-reactivity of three commercial in-house immunoassays for detection of Dirofilaria immitis antigens with Spirocerca lupi in dogs with benign esophageal spirocercosis.

The nematode Spirocerca lupi infects dogs and is endemic in Israel. It leads to formation of esophageal nodules and neoplasia. Infection is diagnosed by coproscopy, endoscopy and radiography. Dirofilaria immitis causes heartworm disease in dogs, and has a world-wide distribution, but autochthonous infection has never been detected in Israel. Infection is confirmed based on identifying D. immitis microfilariae, in concentrated blood specimens by microscopy (Knott's test or other tests) and serological tests specifically detecting circulating adult worm antigens. In the first part of this study, sera from dogs definitively diagnosed with esophageal spirocercosis by endoscopy were examined using three in-house immunoassays for detecting D. immitis antigen, and were positive in 2/19 (10.5%), 5/35 (14.3%) and 14/48 (29.2%) dogs, in assays 1 to 3, respectively, with no statistical difference between assays (P=0.08). Next, sera from 32 additional dogs with confirmed esophageal spirocercosis, which were confirmed to be negative for D. immitis and Dirofilaria repens DNA using a sensitive high-resolution melt PCR were tested using assay 3 and 8/32 (25%) were positive. These results demonstrate serological cross-reactivity between D. immitis and S. lupi in blood samples of dogs. In areas where the distributions of both nematodes overlap, this cross-reactivity should be considered when dog are screened for heartworm disease.

Spirocercosis in dogs in Israel: A retrospective case-control study (2004-2009).

This case-control retrospective study (years 2004-2009) investigated the epidemiological, clinical, and diagnostic test findings of dogs with esophageal spirocercosis (ES) presented to the Hebrew University Veterinary Teaching Hospital (HUVTH) and coproscopy-positive dogs at the Kimron Veterinary Institute (KVI), Israel. It included 133 dogs with ES and 133 negative controls diagnosed at the hospital, and 343 dogs diagnosed at the KVI. The average incidence of ES at the HUVTH was 22.5/year, and the percentage of spirocercosis cases was stable at both institutions (HUVTH, 0.67-1.23%; KVI, 5-8%). Dogs aged > 5 years old had 100-fold likelihood to be infected compared to dogs aged ≤ 1 year of age (P < 0.001). Mean body weight (P = 0.0004), proportion of Retrievers (P = 0.002) and sporting breed dogs (P = 0.006) were higher, while proportion of toy breeds (P = 0.004) was lower in the ES group compared to the control group. The proportion of cases from Greater Tel-Aviv decreased (P = 0.002), while that of those from Judea and Jerusalem increased (P = 0.01) compared to the 1990 s. Spirocercosis occurred in 22 dogs despite past prophylactic avermectin treatment. Vomiting and regurgitation were the most common clinical signs of ES. Coproscopy was S. lupi-positive in 33/60 dogs (55.0%). The median number of esophageal nodules was two (range 1-8), with a median diameter of 3.5 cm (range 1.0-11.0). Malignant esophageal lesion transformation was confirmed in 29 dogs (22%). Despite preventive attempts, spirocercosis has spread in Israel over time, compared to previous findings, raising questions about the efficacy of the currently accepted prophylactic protocol is incompletely effective.

Serum acute phase protein concentrations in dogs with spirocercosis and their association with esophageal neoplasia - a prospective cohort study.

Spirocerca lupi, the dog esophageal worm, typically induces formation of esophageal nodules, which may transform to sarcoma. Ante mortem discrimination between benign and malignant esophageal masses is challenging. Serum acute phase proteins (APPs) are utilized in diagnosis and prognosis of various canine diseases as markers of inflammation. This study characterized serum APPs concentrations in dogs with benign and malignant esophageal spirocercosis and evaluated their accuracy in differentiating benign from malignant lesions. Seventy-eight client-owned dogs with esophageal spirocercosis were included. Serum C-reactive protein (CRP), haptoglobin, serum-amyloid A (SAA) and albumin concentrations were measured upon diagnosis and follow-up visits, and compared with healthy dogs, and between malignant and benign cases. Haptoglobin, CRP and SAA concentrations were higher, and albumin concentration was lower (P<0.001 for all) in infected dogs compared to healthy controls. Dogs with suspected neoplasia had significantly higher CRP (P=0.011), haptoglobin (P=0.008) and SAA (P=0.05), and lower albumin (P=0.012) concentrations compared to dogs with benign esophageal nodules. APPs moderately discriminated between suspected malignant and benign esophageal disease. None of the dogs with suspected neoplasia had concurrent normal concentrations of all APPs. The present results indicate that canine spirocercosis is characterized by an acute phase reaction, both at presentation and during treatment. When concentrations of all four APPs are within reference range, esophageal malignancy is highly unlikely. Although concentrations of all positive APPs were significantly higher in suspected neoplastic cases compared to benign ones, moderate discriminatory power limits their clinical use. Neither APP was useful to monitor response to treatment.

Development and in vitro characterization of floating sustained-release drug delivery systems of polyphenols.

The aim of this study was to develop and characterize floating stomach-retentive matrix tablets that will deliver polyphenols in a controlled release manner. The tablets were prepared by direct compression. A number of polymers were examined and egg albumin was chosen in light of a better performance in terms of floating behavior and decomposition time. Dissolution studies for three representative polyphenols loaded into a number of formulations were performed using the "f2" factor in order to compare release profiles of different polyphenols and formulations. The release data showed a good fit into the power law equation and zero-order kinetics has been determined for some of the systems. Erosion and textural analysis studies revealed that higher concentration of egg albumin results in a higher gel strength that is less susceptible to erosion, potentially leading to a prolonged delivery time of drug. The ability of egg albumin-based tablets to resist high mechanical forces was also determined, while comparison to cellulose-derived polymers revealed that the latter have a much lower ability to resist the same forces. The developed delivery system has the potential to increase the efficacy of the therapy for various pathological stomach conditions and to improve patient compliance.

Sustained-release antibacterial varnish-coated biopsy needle for reduction of infection rates following prostate biopsy-in vitro model.

BACKGROUND AND PURPOSE: During the past decade, the incidence of severe infections after transrectal ultrasonography-guided prostate biopsy has increased. Antibacterial sustained-release varnish has been shown to reduce bacterial infections. This varnish has yet to be tested in the field of urology. We undertook an in vitro study to assess the possibility of reducing infection rates after prostate biopsy by coating the needle with a novel sustained, yet rapid release chlorhexidine varnish (SRV-CHX) specifically modified for prostate biopsy. MATERIALS AND METHODS: A model simulating the microbiologic environment of a prostate biopsy was developed. The model consisted of two layers of agar, of which the first represented the rectum and was preinfected with Escherichia coli. The second layer was sterile and represented tissue interposed between the rectum and prostate. SRV-CHX-coated biopsy needles were inserted 12 times through the two layers, into the third agar layer representing the prostate. Infection transmission was determined by assessing bacterial growth at inoculation sites within the agar plate representing the prostate. Bacterial growth inhibition was measured as an inhibition zone on the contaminated agar. RESULTS: Testing the antibacterial effect of the SRV-CHX-coated needle, we found a substantial reduction of infection transmission as well as sustained inhibition of bacterial growth compared with control needles. CONCLUSIONS: Needles coated with SRV-CHX offer a new strategy in infection control after prostate biopsy. A new strategy of SRV-coated prostate biopsy needles supplemented with various antibacterial agents, combined with prophylactic oral antibiotics should result in decreased infection rates after prostate biopsies. Further in vitro studies are needed to formulate the SRV with an optimal antibacterial agent.

Serum alkaline phosphatase activity is not a marker for neoplastic transformation of esophageal nodules in canine spirocercosis.

BACKGROUND: Spirocerca lupi is a nematode of Canidae that matures within the esophageal wall to form fibroblastic nodules with potential for malignant transformation. Diagnosis is based on histopathologic examination, but false-negative results may be obtained from samples collected by endoscopy. Serum alkaline phosphatase (ALP) activity, frequently increased in hepatobiliary disease, is also increased in a variety of neoplastic conditions in dogs, including appendicular osteosarcoma, and has also been reported to be increased in dogs with spirocercosis. OBJECTIVE: The aim of this study was to evaluate serum ALP activity as a marker for malignant transformation of esophageal nodules in S. lupi-infected dogs. METHODS: In this retrospective study, medical records of dogs diagnosed with spirocercosis from 1991 to 2008 were reviewed, and serum ALP activity determined at presentation was compared between dogs with nonneoplastic and neoplastic nodules. Owing to use of multiple analyzers, ratios of ALP activity to the upper reference interval for ALP were calculated and compared. RESULTS: Median ALP activity ratios were 0.65 (0.07-4.00) and 0.86 (0.10-3.40) for dogs with nonneoplastic (n=88) and neoplastic (n=32) nodules, respectively, with no significant difference (P=.18) and substantial overlap between groups. Tumors included osteosarcoma (15 dogs), fibrosarcoma (15 dogs), and anaplastic sarcoma (2 dogs); there was no difference in ALP activity between the dogs with osteosarcoma and fibrosarcoma. CONCLUSION: ALP is a poor marker of malignant transformation in canine spirocercosis.

Urethral obstruction in cats: predisposing factors, clinical, clinicopathological characteristics and prognosis.

Feline lower urinary tract diseases in general, and urethral obstruction (UO) in particular, are common clinical conditions in cats. The aims of this study were to identify risk factors for UO, to characterise clinical and clinicopathological signs, outcome and recurrence, as well as risk factors for mortality and recurrence. Eighty-two cats with UO were compared to 82 sex and time matched controls. The mean age of cats with UO was significantly lower compared to controls, while the mean body weight was higher. The proportion of indoors-outdoors cats was significantly lower in the study group compared to the control group, and the proportion of cats consuming only dry food was higher. Overall mortality was 8.5%. Ionised calcium was significantly higher in survivors compared to non-survivors, and the prevalence of hypocalcaemia was lower. Recurrence in 6 months and 2 years were 22% and 24%, respectively. Cats with recurrence had significantly lower urine pH at presentation.

Effect of mode of administration on guaifenesin pharmacokinetics and expectorant action in the rat model.

AIM: Guaifenesin is a very commonly used and prescribed oral expectorant drug. However, its mechanism of action is not completely elucidated and the available information is limited. The purpose was to evaluate whether guaifenesin action on respiratory tract secretion is mediated through a reflex stimulation of the gastric mucosa or by the systemic exposure due to the absorption of the drug to the blood circulation. METHODS: Guaifenesin was administered to rats by various routes: intravenous bolus, oral gavage, and gastric, jejunal or cecal infusions (through surgically implanted catheters). Phenol red respiratory tract secretion (after intraperitoneal or intravenous injection) was used as a marker for degree of expectorant action. Administration of saline by gavage was used as control. RESULTS: Respiratory secretion following oral bolus was approximately 2-fold higher (p<0.05) than that of control. Following IV administration the increase of respiratory secretion did not occur despite the fact that systemic exposure to guaifenesin was 1.5-fold higher than following oral administration. The abdominal surgery was found to eliminate the effect of guaifenesin although it did not change systemic absorption. Guaifenesin was equally absorbed from all parts of the gastrointestinal tract. CONCLUSIONS: It was demonstrated that expectorant action of guaifenesin is mediated by stimulation of the gastrointestinal tract and not by the systemic exposure to the drug.

Copromicroscopic and molecular assays for the detection of cancer-causing parasitic nematode Spirocerca lupi.

Spirocerca lupi (Nematoda, Spirurida) is a life-threatening parasitic nematode of dogs that is presently emerging in several countries. Nonetheless, canine spirocercosis is neglected and underestimated, mainly due to diagnostic limitations inherent to clinico-pathologic, diagnostic imaging and laboratory methodologies. Given the significant benefit of improved diagnosis, the present work evaluated the reliability of a recently described copromicroscopic approach, the FLOTAC technique, as well as a PCR-based assay with that of traditional coproscopic techniques to diagnose S. lupi infection. Ninety-four faecal field samples were collected from two endemic areas (i.e. 29 and 65 from Kenya and Israel, respectively) and processed using different coproscopic examination techniques. In particular, set I (Kenyan samples) comprised the modified flotation with Sheather's sugar solution and merthiolate-iodine-formalin technique, while set II (Israeli samples) comprised a flotation technique with zinc sulphate solution, a modified sugar flotation procedure and the FLOTAC method. All samples were also subjected to a semi-nested PCR protocol specific for a region internal to the mitochondrial cytochrome c oxidase subunit 1 gene of S. lupi. The coproscopic examinations showed low sensitivity and high variability, demonstrating the unreliability of the conventional methods for detecting S. lupi eggs. Nonetheless, the FLOTAC technique scored the highest number of positives and significantly higher number of S. lupi eggs per microscopic field compared to the other coproscopic methods. Additionally, of the coproscopically negative samples, 9 (45%) Kenyan and 21 (38.2%) Israeli samples scored molecularly positive using the PCR-based approach. The potential implications and perspectives for canine spirocercosis of these coproscopic and molecular diagnostic methodologies evaluated herein are discussed.

Transient megaoesophagus and oesophagitis following diaphragmatic rupture repair in a cat.

A 6-month-old domestic shorthair female cat was presented with suspected diaphragmatic hernia (DH) that was later confirmed by thoracic radiography. The cat underwent exploratory celiotomy with a diaphragmatic rupture (DR) repair and recovered. Six days later, it was represented with vomiting and anorexia. Megaoesophagus (MO) and gastric dilatation were diagnosed by contrast radiography. A second celiotomy revealed no abnormalities and gastropexy was performed. Endoscopy demonstrated MO, oesophagitis and gastro-oesophageal reflux. MO persisted for several weeks and was an unexpected complication as no association between DR (or DH) and MO has never been described in the veterinary literature. The cat was treated medically with aggressive prokinetic and antacid therapy along with prolonged temporary oesophageal diversion (percutaneous endoscopic gastrostomy tube) with an excellent outcome.

Oesophageal sarcomas in dogs: histological and clinical evaluation.

A histological grading system of oesophageal sarcomas has not been established. Thirty-two cases of oesophageal sarcomas have been reviewed for tumour characteristics and clinical outcome. Nineteen dogs underwent surgical intervention to remove oesophageal tumours; ten of them survived (median 278 days). Primary tumour types included osteosarcoma (47%), osteosarcoma with tumour giant cells (7%), fibroblastic osteosarcoma (13%), chondroblastic osteosarcoma (7%) fibrosarcoma (23%) and undifferentiated sarcoma (3%). Histological grade evaluation revealed 33% grade 1 sarcoma, 50% grade 2 and 17% grade 3. No correlation could be found between survival and signalment, duration of clinical signs, tumour type, tumour grade and chemotherapy. Chemotherapy was found to reduce lung metastases' histological scores in three cases (P=0.0007). Surgery seems to be the treatment of choice but the effect of chemotherapy warrants further investigation. Additional research of cases should be performed in order to further define prognostic factors of oesophageal sarcomas.

Chemotherapeutic treatment of xenograft Spirocerca lupi-associated sarcoma in a murine model.

To date, data are not available concerning the effectiveness of chemotherapy in the treatment of Spirocerca lupi-associated esophageal sarcomas. In the present study, we compared the effectiveness of 4 chemotherapeutic agents against S. lupi-associated osteosarcoma, using a xenograft murine model created in our lab. Samples of xenografted osteosarcoma were inoculated subcutaneously into 5 groups (n = 10 each) of 6-wk-old male and female NOD/SCID mice. Tumor-bearing mice were divided into treatment and control groups. The treatment groups were injected with either pegylated liposomal doxorubicin (6 mg/kg, intravenously, n = 9), doxorubicin (6 mg/kg, intravenously, n = 8), carboplatin (60 mg/kg, intraperitoneally, repeated twice at 1-wk intervals for a total of 2 doses, n = 9), or cisplatin (6 mg/kg, intraperitoneally, n = 8). The control group was injected with buffered saline (n = 9). Tumor size was determined by caliper measurements. Compared with the control group, the pegylated liposomal doxorubicin- and doxorubicin-treated groups, but not the carboplatin or cisplatin groups, showed significant inhibition of tumor growth. Our results indicate that doxorubicin-based drugs are effective against S. lupi-associated sarcomas in a mouse xenograft model. Because it is less toxic than doxorubicin, pegylated liposomal doxorubicin is likely the drug of choice for treatment of S. lupi-associated sarcomas. We suggest that combination of doxorubicin or its pegylated form with surgical excision will improve the prognosis of dogs with this disease.

Spirocercosis-associated pyothorax in dogs.

The records of five dogs diagnosed with spirocercosis-associated pyothorax were retrospectively reviewed. On admission, the most common clinical findings were weakness, anorexia, depression, dyspnoea and fever. All dogs had typical oesophageal Spirocerca lupi granulomas. Contrast thoracic radiography revealed contrast material leakage from the oesophagus into the pleural cavity in two dogs only. All dogs were medically treated with broad-spectrum antibiotics, repeated chest tube drainage and thoracic lavage, and four of the dogs were treated with doramectin. One dog was euthanased due to a concurrent non-related disease at the owners request while the remaining four survived. S. lupi infection should be included in the differential diagnoses of canine pyothorax in endemic areas. Conservative medical management of pyothorax with antihelmintic medication was shown to be effective in the treatment of S. lupi-associated pyothorax.

Murine xenograft model of Spirocerca lupi-associated sarcoma.

Nodular masses and granulomas of the esophagus are among the most frequent lesions caused by Spirocerca lupi, a nematode parasite of dogs, and neoplastic transformation of these granulomas to osteosarcoma or fibrosarcoma has been described. In this study, we developed a xenograft murine model of S. lupi-associated sarcoma. Samples of esophageal fibrosarcoma and osteosarcomas were excised from three dogs diagnosed with spirocercosis. These sarcomas were inoculated into three groups of 6-week-old NOD/SCID mice to create three tumor lines of S. lupi-associated sarcomas. Mice in all groups developed tumors after inoculation, and the cell lines could be further propagated as second-generation xenografts. We successfully established xenograft murine models of three different lines of S. lupi-associated sarcoma that offer readily available sources of these tumors for further experiments. This resource will facilitate studies on the malignant transformation of the granulomas, establishment of efficient chemotherapy and radiotherapy regimens, and identification of diagnostic molecular markers.

Spirocercosis-associated esophageal sarcomas in dogs. A retrospective study of 17 cases (1997-2003).

Seventeen client-owned dogs diagnosed with spirocercosis-associated esophageal sarcomas were retrospectively reviewed. The most common clinical signs noticed were vomiting and/or regurgitation (94%), lethargy and depression (59%), pyrexia and anorexia (41% each). Leukocytosis (82%) and microcytic hypochromic anemia (30%) were the most common hematological abnormalities. Caudal thoracic masses were demonstrated on survey radiographs of 13/15 of the dogs and thoracic spondylitis was detected in 12/15 dogs. Spirocerca lupi eggs were detected in 2/8 patients and worms were demonstrated on 1/11 at necropsy. Ten cases underwent surgical attempt to remove the tumors. In six of them partial esophagectomy (PE) was performed and all of them survived the immediate postoperative hospitalization. Five of the cases that underwent PE also received chemotherapy after surgery (doxorubicin (Adriamycin, Upjohn)) with an average survival time of 267 days. The histopathological results of the esophageal tumors were osteosarcoma (9), fibrosarcoma (5) and undifferentiated sarcoma (1). In areas endemic to spirocercosis, regurgitation or vomiting in dogs and microcytic hypochromic anemia and neutrophilia warrant ruling out esophageal sarcomas. Proper surgical treatment could prolong the dogs' lifespan for months, and improve their quality of life.

Evaluation of doramectin for the treatment of experimental canine spirocercosis.

The nematode Spirocerca lupi is primarily a parasite of dogs, which causes typical lesions of esophageal nodular granulomas, aortic aneurysms and spondylitis. In order to evaluate the therapeutic effect of doramectin on experimental canine spirocercosis, seven beagle dogs experimentally infected with 40 infectious S. lupi larvae (L(3)) were treated with doramectin. Treatment was commenced following endoscopic visualization of esophageal granulomas, and typical S. lupi eggs were detected in the feces. The treatment protocol included six treatments of doramectin (400 microg/kg subcutaneously) at 2 weeks intervals, followed by monthly injections until the disappearance of the esophageal granulomas or the end of the study (768 days post-inoculation). Eggs could not be found on fecal examinations 3-10 days after the first or second doramectin treatment. In addition, a gradual decrease in size of granulomas was noticed in all seven dogs during the course of the study. Esophageal granulomas had completely resolved in six of the seven dogs between day 35 and day 544 post-initial doramectin treatment, by day 35 in one dog (after three treatments), by day 43 in two dogs (after four treatments), by day 98 in one dog (after seven treatments), by day 460 in one dog (after 18 treatments) and by day 544 in another dog (after 21 treatments). In one dog, remnants of S. lupi granulomas could still be seen 544 days post-initiation of treatment with doramectin. Multiple subcutaneous injections of doramectin (400 microg/kg) were shown to be effective and safe in the treatment of canine spirocercosis.

Canine spirocercosis: clinical, diagnostic, pathologic, and epidemiologic characteristics.

The nematode Spirocerca lupi is a parasite of dogs with beetles of several species serving as intermediate hosts. The medical records of 50 dogs diagnosed with spirocercosis at the Hebrew University Veterinary Teaching Hospital (HUVTH) in Israel during 1991-1999 were retrospectively reviewed and compared to a control group (n=100). There was a seven-fold increase in the annual number of dogs diagnosed with spirocercosis during these years while the hospital caseload increased by 80%, indicating an emerging outbreak of this infection. Dogs from the greater Tel Aviv area were at the highest risk of being diagnosed with spirocercosis with 74% of the cases originating from this region compared to only 17% of the controls. The disease appeared to have a primarily urban pattern of distribution with a significantly higher percentage (P=0.025) of dogs from cities versus rural areas, as compared to the control group. Sixty-two percent of the cases were diagnosed during the colder months of December through April. The median age of infected dogs was 5 years, with dogs 1 year old or younger at the lowest risk of being diagnosed with spirocercosis. Large breeds were at a higher risk of infection in comparison to small breeds and the Labrador Retriever was significantly over represented (P=0.027) in the study group compared to the control population. The most common signs were vomiting or regurgitation (60%), pyrexia (24%), weakness (22%), respiratory abnormalities (20%), anorexia (18%), melena (18%) and paraparesis (14%). A caudal esophageal mass was identified by radiography in 53% of the dogs and spondylitis of the thoracic vertebrae in 33%. Fecal flotation was positive for S. lupi eggs in 80% of the dogs, and endoscopy was found to be the most sensitive diagnostic procedure and allowed diagnosis in 100% of the examined dogs. Fifty-three percent of the dogs were anemic and creatine kinase (CK) activities were elevated in 54%. Necropsy of 14 dogs revealed esophageal or gastric granulomas in 13 dogs, and an esophageal osteosarcoma in a single animal. Aortic aneurysms were found in six (43%) dogs. Out of 24, 15 dogs (63%) for which follow-up information was available died or were euthanized within 1 month of admission. The case-fatality rate decreased toward the end of the study period when improved therapy with avermectins became available.