Acute phase proteins, interleukin-6, tumor necrosis factor, nitric oxide and oxidative stress markers in horses with cutaneous habronemosis under field condition.

Habronemosis is a common parasitic disease of horses worldwide. In order to investigate how haptoglobin (Hp), serum amyloid A (SAA), oxidative stress markers, nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor (TNF- α), varies in cutaneous habronemosis, 30 horses with the clinical disease and 20 clinically healthy horses were included in the current study. The serum levels of Hp, SAA, and proinflammatory cytokines (IL-6 and TNF-α), NO, malondialdehyde (MDA), super oxide dismutase (SOD), glutathione (GSH), and total antioxidant capacity (TAC) were determined in horses before and after two weeks of treatment. The serum levels of Hp, SAA, IL-6, TNF-α and MDA were significantly elevated in infected horses as compared to the controls. Alternately, the serum levels of SOD, GSH, TAC and NO, were recorded low in infected horses as compared to the controls. All tested markers resumed the same levels after treatment as in control group. The Hp, SAA, IL-6, TNF-α, and MDA exhibited a high degree of clinical accuracy of the cases diagnosis. The area under the curve (AUC) for acute phase proteins (SAA, Hp), IL-6, TNF-α, and MDA was 0.87, 0.94, 0.96, 0.96 and 1.0, respectively. These findings showed that Hp, SAA, IL-6, TNF-α, and MDA may be supportive in the diagnosis of cutaneous habronemosis in horses and, simultaneously, they can also be used to monitor the progress of the treatment.

The endocannabinoid system in canine Steroid-Responsive Meningitis-Arteritis and Intraspinal Spirocercosis.

Endocannabinoids (ECs) are involved in immunomodulation, neuroprotection and control of inflammation in the central nervous system (CNS). Activation of cannabinoid type 2 receptors (CB2) is known to diminish the release of pro-inflammatory factors and enhance the secretion of anti-inflammatory cytokines. Furthermore, the endocannabinoid 2-arachidonoyl glycerol (2-AG) has been proved to induce the migration of eosinophils in a CB2 receptor-dependent manner in peripheral blood and activate neutrophils independent of CB activation in humans. The aim of the current study was to investigate the influence of the endocannabinoid system in two different CNS inflammatory diseases of the dog, i.e. Steroid-Responsive Meningitis-Arteritis (SRMA) and Intraspinal Spirocercosis (IS). The two main endocannabinoids, anandamide (AEA) and 2-AG, were quantified by mass spectrometry in CSF and serum samples of dogs affected with Steroid- Responsive Meningitis-Arteritis in the acute phase (SRMA A), SRMA under treatment with prednisolone (SRMA Tr), intraspinal Spirocercosis and healthy dogs. Moreover, expression of the CB2 receptor was evaluated in inflammatory lesions of SRMA and IS and compared to healthy controls using immunohistochemistry (IHC). Dogs with SRMA A showed significantly higher concentrations of total AG and AEA in serum in comparison to healthy controls and in CSF compared to SRMA Tr (p<0.05). Furthermore, dogs with IS displayed the highest ECs concentrations in CSF, being significantly higher than in CSF samples of dogs with SRMA A (p<0.05). CSF samples that demonstrated an eosinophilic pleocytosis had the highest levels of ECs, exceeding those with neutrophilic pleocytosis, suggesting that ECs have a major effect on migration of eosinophils in the CSF. Furthermore, CB2 receptor expression was found in glial cells in the spinal cord of healthy dogs, whereas in dogs with SRMA and IS, CB2 was strongly expressed not only in glial cells but also on the cellular surface of infiltrating leukocytes (i.e. neutrophils, eosinophils, lymphocytes, plasma cells, and macrophages) at lesion sites. The present study revealed an upregulated endocannabinoid system in dogs with inflammatory CNS diseases, highlighting the endocannabinoid system as a potential target for treatment of inflammatory CNS diseases.

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.

Vascular endothelial growth factor concentrations in dogs with spirocercosis.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent proangiogenic factor associated with tumor development. Spirocerca lupi is a nematode of canids that induces an esophageal nodule that progresses to a sarcoma in 25% of cases. Determination of neoplastic transformation is challenging and usually based on endoscopy-guided biopsies under general anesthesia, an expensive procedure that often yields nondiagnostic, necrotic samples. HYPOTHESIS: Circulatory VEGF concentrations are increased in dogs with neoplastic spirocercosis and can distinguish between dogs with neoplastic and nonneoplastic disease. ANIMALS: A total of 24 client-owned dogs, 9 nonneoplastic, 9 neoplastic, and 6 controls. METHODS: Case-control study. Plasma and serum VEGF concentrations at the time of diagnosis were compared with those of healthy controls. Measurement of VEGF was performed using a canine-specific ELISA. Kruskal-Wallis and Dunn's tests were used for statistical analysis with significance set at P < .05. RESULTS: The median plasma VEGF concentrations of dogs with neoplastic spirocercosis were 629 pg/mL (range, 282-2,366) higher than both the nonneoplastic (<39.5 pg/mL; range, <39.5-716) and control dogs (<39.5 pg/mL; all values, <39.5; P = .0003). The median serum VEGF concentration of the neoplastic dogs was 69 pg/mL (range, <39.5-212) higher than the nonneoplastic (<39.5 pg/mL; range, <39.5-44.13) and control dogs (<39.5 pg/mL; all values, <39.5; P = .001). CONCLUSIONS AND CLINICAL IMPORTANCE: Both plasma and serum VEGF concentrations can be used to differentiate nonneoplastic and neoplastic spirocercosis. The role of VEGF in neoplastic transformation of S. lupi-induced nodules and the potential utility of anti-VEGF drugs in spirocercosis-induced sarcoma warrant further investigation.

Hypovitaminosis D in dogs with spirocercosis.

BACKGROUND: Spirocercosis in dogs is characterized by esophageal nodules that can undergo neoplastic transformation. Hypovitaminosis D has been associated with neoplasia formation. We hypothesized hypovitaminosis D in neoplastic spirocercosis and that it could be a risk factor for neoplastic transformation. OBJECTIVE: To measure and compare vitamin D status, assessed by serum 25-hydroxyvitamin D [25(OH)D] concentrations in non-neoplastic (n = 25) and neoplastic (n = 26) spirocercosis client-owned dogs and healthy dogs (n = 24). ANIMALS: Twenty-five non-neoplastic dogs, 26 neoplastic dogs, and 24 healthy dogs. METHODS: Fifty-one dogs were randomly selected from 119 dogs diagnosed with spirocercosis presenting to our hospital, and further divided into non-neoplastic or neoplastic groups. Exclusion criteria included dogs less than 1 year old, with concurrent diseases, received corticosteroids, or treated prophylactically for spirocercosis. Serum 25(OH)D concentration was measured by high-performance liquid chromatography. Spirocercosis dogs' appetites were graded and compared. RESULTS: Serum 25(OH)D concentrations were significantly different among all groups (P < .001). 25-Hydroxyvitamin D concentrations were significantly lower in neoplastic group (median 30.7 nmol/L [range 14.7-62.2]) compared to non-neoplastic (median 52.7 nmol/L [range 19.1-129.7, P < .05]) and healthy groups (median 74.6 nmol/L [range 37.4-130.5, P < .005]). 25-hydroxyvitamin D concentrations were significantly lower in non-neoplastic spirocercosis dogs compared to healthy ones (P < .05). Neoplastic and non-neoplastic spirocercosis dogs had similar appetite scores (P = 1.0). 25-Hydroxyvitamin D concentrations were not significantly different between dogs with normal (P = .087) and abnormal (P = .125) appetites within neoplastic and non-neoplastic spirocercosis groups. CONCLUSIONS AND CLINICAL IMPORTANCE: Further studies are warranted to determine potential use of vitamin D treatment in spirocercosis and explore role of hypovitaminosis D in pathogenesis of malignant transformation.

Serum C-reactive protein concentration in benign and malignant canine spirocercosis.

BACKGROUND: Spirocerca lupi is a nematode of canids that forms a nodule in the esophagus that can undergo neoplastic transformation. C-reactive protein (CRP) is a major acute phase protein in the dog that has been used for treatment, monitoring, and prognostication in inflammatory and neoplastic disease. HYPOTHESIS/OBJECTIVES: The objectives of this study were to determine if serum CRP concentration (1) is increased in canine spirocercosis, (2) can be used to determine neoplastic transformation, and (3) can be used to monitor response to treatment in benign spirocercosis. ANIMALS: Forty-two dogs naturally infected with S. lupi and 21 control dogs. METHODS: A retrospective study was performed. The infected cases were divided into benign (n = 28) or malignant (n = 14) spirocercosis. CRP was performed on all of the spirocercosis and control cases at presentation. Statistical analysis was done by the one-way analysis of variance and Student's t-test. RESULTS: The mean CRP concentration in the benign cases was 60.4 ± 48.0 mg/L and that of the malignant cases was 76.5 ± 44.8 mg/L; both values were significantly higher (P < .001) than those of the control group where the mean was 13.4 ± 17.9 mg/L. The mean CRP concentration for the convalescent sera in the benign group was lower than the pretreatment concentrations (P= .01). CONCLUSION AND CLINICAL IMPORTANCE: CRP cannot be used to differentiate between benign and malignant spirocercosis. There is a decrease in CRP concentration in dogs with benign spirocercosis once treatment has commenced. Serial CRP measurement can be used to monitor response to treatment in benign spirocercosis.

Protein profile analysis from advanced third-stage larvae (AdvL3) and adult worms of Gnathostoma binucleatum (Nematoda: Spirurida).

Proteins from crude extracts of advanced third-stage larvae and adult Gnathostoma binucleatum nematode worms showed protein profiles in SDS-PAGE analysis similar to Echinococcus granulosus, Trichinella spiralis, Dipylidium caninum, Ancylostoma caninum, Ascaris lumbricoides and Toxocara canis. The immunoblot analysis of the human serum infected or suspected to be infected with G. binucleatum using the total larvae extract recognized the 40, 60, 80 and 115 kDa proteins and using the total adult worm extract recognized only the 80 and 115 kDa proteins. However, the 115 kDa protein showed cross-reactions with A. caninum, A. lumbricoides, T. canis and D. caninum with human serum positive to gnathostomosis, while the 40 kDa protein was only recognized with the G. binucleatum total larvae extract. The results obtained suggest that the use of antigens from the advanced third-stage larvae of the parasite were best recognized for immunodiagnosis of gnathostomosis.

Diagnostic values of IgG4 in human gnathostomiasis.

The diagnostic values of immunoglobulin G subclass antibodies from patients with gnathostomiasis were assessed by immunoblot technique. Antigen was prepared from crude extracts of Gnathostoma spinigerum advanced third-stage larvae obtained from naturally infected eels. The sera were obtained from 14 parasite-confirmed gnathostomiasis cases, 63 patients with other helminthic infections and 13 healthy controls. Nine prominent IgG4 reactive bands appeared with molecular weights of 94, 51, 47, 43, 38, 24, 21, 20 and 15 kDa. The diagnostic sensitivity of each of the nine reactive bands ranged from 100% to 64.3% in 14 parasite-confirmed gnathostomiasis cases. All (100%) confirmed cases recognized the 21 kDa antigenic band, but not other helminthic infections or parasite-free control. Recognition of 21 kDa antigen in G. spinigerum advanced third-stage larvae crude extracts is the most specific diagnostic marker for human gnathostomiasis, with 100% sensitivity and specificity. The 20 and 24 kDa protein bands were additional diagnostic bands for confirming diagnosis of infection where the 21 kDa band was faint. No specific binding of IgG1, IgG2, or IgG3 antibodies was observed in any sera from confirmed gnathostomiasis cases.

Double-dose ivermectin vs albendazole for the treatment of gnathostomiasis.

A comparative study was performed for the treatment of gnathostomiasis patients with ivermectin 0.2 mg/kg for 2 days in 15 patients vs albendazole 400 mg twice daily for 21 days in 14 patients. The ivermectin and albendazole gave cure rates of 100% and 78.5%, respectively, however the difference was not statistically significant between the two drugs (Fisher's exact, p=0.0996). One year after treatment, the patients who had no migratory swellings and a drop in ELISA titers or a negative immunoblot test were considered to be cured. The side effect of ivermectin for two days was dizziness. The side effects of albendazole were nausea, dizziness, and an increased alkaline phosphatase.

Gnathostomiasis possibly caused by Gnathostoma malaysiae.

Gnathostomiasis is rarely reported in travelers, although the disease remains a major public health problem in Southeast Asia. A creeping eruption and Quincke's edema (slowly migrating erythema with pruritus) appeared in two Japanese men who had eaten raw freshwater shrimp in Myanmar. A Gnathostoma larva was found in subcutaneous tissue from one of the men. Four species causing human gnathostomiasis, G. hispidum, G. doloresi, G. nipponicum and G. spinigerum, can be distinguished based on the number of nuclei in intestinal epithelial cells of infected larvae, in cross-section. In G. hispidum, only a single large nucleus is found. Morphologically, our larva was initially identified as G. hispidum. However, since the number of epithelial cells was greater and the body width was larger than those of a "large-type" 3rd-stage larva of G. hispidum, the larva was then identified as a 3rd-stage larva of G. malaysiae, Miyazaki and Dun, 1965, as reported by Setasuban et al, (1991). Since no human cases caused by this species of Gnathostoma have previously been encountered, this appears to be the first report of gnathostomiasis due to G. malaysiae.

Physiological status of wild European eels (Anguilla anguilla) infected with the parasitic nematode, Anguillicola crassus.

The effect of the parasitic swimbladder nematode, Anguillicola crassus, on the physiological status of wild European eels (Anguilla anguilla) was investigated during an 18 month survey (February 1995 until September 1996), and compared with that of wild uninfected eels collected over the same time-period. Despite the occurrence of up to 15 blood-feeding adults in the swimbladder lumen of the infected eels and as many as 25 additional larvae in the swimbladder wall, there were no major differences in hormonal, metabolic or osmoregulatory status of the 2 groups of eels. Wild European eels appear to adapt to chronic parasitism with Anguillicola crassus. The possible adverse effects of additional simultaneous stressors are discussed.