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1.
Vet Parasitol ; 206(1-2): 55-9, 2014 Nov 15.
Article in English | MEDLINE | ID: mdl-25224786

ABSTRACT

Adulticide treatment of dogs with canine heartworm disease causes the death of the adult Dirofilaria immitis lodged in the vascular system of the host. During the death of the worms, pulmonary thromboembolisms (PTE), pulmonary inflammation, congestive heart failure, or renal disease are possible consequences. The aim of this study was to evaluate cardiopulmonary biomarkers and renal parameters during adulticide treatment of canine heartworm to compare the classic two-injection treatment protocol versus the American Heartworm Society (AHS) recommended protocol. Fourteen heartworm-infected dogs with high parasite burdens were divided in three groups and allocated to adulticide protocols as follows: Group A (n=5) was treated with the classic two-injection protocol; group B (n=5) was treated using the AHS recommended protocol, and group C (n=4) was treated as for group B but received diminishing anti-inflammatory doses of prednisone. To assess cardiorespiratory status, cardiac troponin I (cTnI), myoglobin, and D-dimer were measured. Renal function was evaluated by measuring urea, creatinine, and urine protein:creatinine (UP:C). Serum and urine samples were collected day 0 (day of diagnosis), 7 and 14 days after the first dose of adulticide, and 1 month after the last adulticide injection. Dogs that received classic treatment presented pathologic concentrations of D-dimer more frequently and showed higher average D-dimer levels, which may indicate the presence of more severe PTE. Group C showed the highest levels of D-dimer during treatment, which may be due to an exacerbation of PTE caused by the administration of prednisone. CTnI and myoglobin values remained above reference values in all groups during the study but reached the lowest values 1 month after the last injection. Levels of urea and creatinine were within normal ranges in all groups, and 28.5% of the dogs were proteinuric on day 0, progressing to better UP:C values at the end of the treatment, except in group A. The results of this study justify the treatment of canine heartworm disease using the AHS recommended adulticide guidelines and recommends re-evaluation of the role of glucocorticosteroids in the prevention and treatment of PTE.


Subject(s)
Clinical Protocols/standards , Dirofilariasis/blood , Dirofilariasis/drug therapy , Dog Diseases/drug therapy , Filaricides/therapeutic use , Animals , Biomarkers/blood , Dirofilaria immitis , Dog Diseases/blood , Dog Diseases/parasitology , Dogs , Practice Guidelines as Topic/standards , United States
2.
Vet Parasitol ; 206(1-2): 43-7, 2014 Nov 15.
Article in English | MEDLINE | ID: mdl-25224789

ABSTRACT

The prognosis and success of adulticide treatment depends on the baseline severity of infection with Dirofilaria immitis in dogs and can influence the therapeutic protocol to choose. A study was conducted to assess the utility of the cardiopulmonary biomarkers N-terminal of the prohormone brain natriuretic peptide (NT-proBNP), cardiac troponin I (cTnI), myoglobin, and D-dimer, as well as the biomarker of inflammation C-reactive protein (CRP) to evaluate the severity of canine heartworm disease in different stages. Serum samples were collected from 20 heartworm-infected dogs for measurement of these biomarkers. Dogs were assigned to four groups (n=5/group) according to the severity of the disease: Class I and Class II dogs showed concentrations of biomarkers generally within normal ranges for healthy dogs, except CRP in dogs classified as Class II, which showed values slightly above laboratory reference values (19.46±5.69mg/l). Dogs of Class III showed elevated levels of NT-proBNP (1220.12±465.18pmol/l) (p<0.05), cTnI (0.99±0.39ng/ml), CRP (33.98±7.99mg/l) (p<0.05), and 40% of these dogs (2/5) presented pathological values of D-dimer and myoglobin (p<0.05). Dogs of Class IV (caval syndrome) presented elevated levels of NT-proBNP (>2530.8pmol/l), cTnI (1.99±0.32ng/ml), and CRP (80.24±47.69mg/l); all Class IV dogs showed pathological elevations of D-dimer (0.81±0.46ng/ml), and 60% (3/5) of these dogs showed pathological elevations of myoglobin (significant elevations [p<0.05] for all biomarkers with respect to reference values and values for dogs in Classes I, II, and III). In canine heartworm disease, chronic presence of D. immitis causes a proliferative endoarteritis, thromboembolisms, pulmonary hypertension, and right-sided congestive heart failure. As the severity of the disease worsens, serum values of cTnI, myoglobin, and NT-proBNP increase, indicating significant cardiac damage. The finding of pathological concentrations of D-dimer suggests the presence of thromboembolism and/or disseminated intravascular coagulation. CRP increases according to the severity of the disease, indicating inflammatory processes that could contribute to the progression of the disease. These preliminary results demonstrate the utility of cardiopulmonary and inflammation biomarkers to assist in the establishment of the severity of canine heartworm disease.


Subject(s)
Biomarkers/blood , Dirofilariasis/pathology , Dog Diseases/pathology , Animals , Dirofilaria immitis/physiology , Dirofilariasis/blood , Dirofilariasis/physiopathology , Dog Diseases/blood , Dog Diseases/physiopathology , Dogs , Inflammation/blood , Severity of Illness Index
3.
Vet Parasitol ; 197(1-2): 244-50, 2013 Oct 18.
Article in English | MEDLINE | ID: mdl-23759483

ABSTRACT

Heartworm disease (Dirofilaria immitis) is a parasitic disease of dogs and other carnivores, characterized by the presence of adult worms in the pulmonary arteries and right ventricle, leading to pulmonary hypertension which may progress to congestive heart failure. Cardiac biomarkers are biological parameters that can be objectively measured as indicators of pathological processes, or to assess the response to therapeutic interventions. To evaluate the myocardial damage during the adulticide treatment in 15 heartworm-infected dogs with ivermectin, doxycycline and melarsomine, measurements of cardiac troponin I (cTnI), myoglobin, MB isoenzyme of creatine kinase (CK-MB) and aspartate aminotransferase (AST) were carried out on days 0, 60 and afterwards weekly on days 67, 75, 82, 91, 106, 113 and 120. Dogs were divided by low parasite burden (n=9) and high parasite burden (n=6). On day 0, dogs with high worm burden showed increased cTnI concentrations (3.62 ± 4.78 ng/ml) while dogs with low worm burden had concentrations similar to those of healthy dogs (0.78 ± 0.22 ng/ml), CK-MB concentrations were increased only in dogs with high parasite burden as well (54.4 ± 54.2U/l) and 26.6% (4/15) of the dogs showed pathological concentrations of myoglobin. On day 91, most dogs showed pathological concentrations of myoglobin, CK-MB and AST, probably due to the myositis associated to the intramuscular injection of melarsomine. The rest of the measurements made in the study, the biomarkers concentrations were within normal values, except for cTnI in dogs with high parasite burden, which remained above reference concentrations for healthy dogs during all the study. The evaluation of cardiac biomarkers seems to be a helpful test in the assessment of the myocardium in dogs with heartworm disease during the adulticide treatment.


Subject(s)
Creatine Kinase, MB Form/blood , Dirofilaria immitis , Dirofilariasis/drug therapy , Dog Diseases/drug therapy , Myoglobin/blood , Troponin/blood , Animals , Anti-Bacterial Agents/administration & dosage , Anti-Bacterial Agents/therapeutic use , Arsenicals/administration & dosage , Arsenicals/therapeutic use , Biomarkers/blood , Biomarkers/metabolism , Creatine Kinase, MB Form/metabolism , Dirofilariasis/metabolism , Dirofilariasis/pathology , Dog Diseases/metabolism , Dogs , Doxycycline/administration & dosage , Doxycycline/therapeutic use , Drug Administration Schedule , Drug Therapy, Combination/veterinary , Filaricides/administration & dosage , Filaricides/therapeutic use , Gene Expression Regulation , Ivermectin/administration & dosage , Ivermectin/therapeutic use , Myoglobin/metabolism , Triazines/administration & dosage , Triazines/therapeutic use , Troponin/metabolism
4.
Vet Parasitol ; 195(1-2): 106-11, 2013 Jul 01.
Article in English | MEDLINE | ID: mdl-23384581

ABSTRACT

Dirofilaria immitis adult worms lodged in the pulmonary arteries are the causative agent of heartworm disease, characterized by endarteritis and obstruction of the blood flow. Dying worms form embolic fragments which trigger thrombosis. Thus, pulmonary thromboembolism is an inevitable consequence of successful adulticide therapy. Clinical signs of pulmonary thromboembolisms are highly variable and non-specific, and its diagnosis is often difficult to obtain. The D-dimer assay is considered the marker of choice for dogs with a suspicion of pulmonary thromboembolism. The aim of the present study was to evaluate the variation of D-dimer and platelets in 15 heartworm-infected dogs during the adulticide treatment with ivermectin, doxycycline and melarsomine. Nine dogs were considered to have a low parasite burden and 6 had a high burden. Measurements were carried out on days 0, 60, and afterwards weekly on days 67, 75, 82, 91, 98, 106, 113, 120, and finally on day 271 after treatment. 40% of dogs showed pathological concentrations of D-dimer and 40% showed slight or mild thrombocytopenia on day 0. The levels of D-dimer decreased by day 60, probably due to the joint action of the ivermectin, doxycycline and exercise restriction. All dogs with high parasite burden showed elevated values of D-dimer on several occasions during the treatment, reaching the highest values the first and second week after the injections of melarsomine. Only 33.3% of the dogs with low parasite burden showed elevated D-dimer levels at some point during the treatment, and all of them showed undetectable levels from the third week after the first injection of melarsomine. On day 271, all dogs showed undetectable levels of D-dimer. There was no correlation between thrombocytopenia and levels of D-dimer during the treatment. The evaluation of serum D-dimer appears to be a supportive test in the assessment and monitoring of pulmonary thromboembolism in dogs with heartworm disease during the adulticide treatment.


Subject(s)
Antifibrinolytic Agents/analysis , Dirofilaria immitis/drug effects , Dirofilariasis/drug therapy , Dog Diseases/drug therapy , Fibrin Fibrinogen Degradation Products/analysis , Pulmonary Embolism/veterinary , Thrombocytopenia/veterinary , Animals , Antiparasitic Agents/therapeutic use , Arsenicals/therapeutic use , Dirofilariasis/parasitology , Dog Diseases/parasitology , Dogs , Doxycycline/therapeutic use , Female , Ivermectin/therapeutic use , Male , Pulmonary Artery/parasitology , Pulmonary Embolism/parasitology , Thrombocytopenia/parasitology , Triazines/therapeutic use
5.
J Anim Physiol Anim Nutr (Berl) ; 96(4): 655-9, 2012 Aug.
Article in English | MEDLINE | ID: mdl-21732995

ABSTRACT

Most pet dogs in developed countries are fed commercial diets. The aim of this study was to evaluate the preferences of owners of overweight dogs when buying commercial pet food. The study was a descriptive observational multi-centre study on a group of 198 owners of urban household dogs. Personal interviews were conducted to examine the owners' opinions with questions rating the importance of certain qualities of prepared dog food. Bivariate analyses for comparisons of absolute means between groups of owners of dogs with excess weight (n = 137) and owners of normal weight dogs (n = 61) were made using the Mann-Whitney U-test. A low price (p < 0.001) and special offers (p = 0.008) of commercial dog food were more important for owners of dogs with excess weight than for owners of normal weight dogs. The quality of ingredients (p = 0.007) and the nutritional composition (p < 0.001) were more important for owners of normal weight dogs than for owners of dogs with excess weight. The veterinarian was the most important source of information on dog nutrition for both groups (83.6% for owners of normal weight dogs and 83.2% for owners of dogs with excess weight) (p = 0.88). The owners of dogs with excess weight had less interest in corrected dog nutrition than owners of normal weight dogs (p < 0.001).


Subject(s)
Animal Feed/analysis , Consumer Behavior , Diet/veterinary , Dog Diseases/diet therapy , Overweight/veterinary , Animal Nutritional Physiological Phenomena , Animals , Consumer Behavior/statistics & numerical data , Data Collection , Dogs , Overweight/diet therapy , Surveys and Questionnaires
6.
Vet Parasitol ; 176(4): 291-4, 2011 Mar 22.
Article in English | MEDLINE | ID: mdl-21310532

ABSTRACT

The island of Gran Canaria is a hyperendemic area for canine dirofilariasis. The aim of the present study was to provide data on Dirofilaria immitis in dogs, cats, and humans on this island in 2010. The data confirms the prevalence in the overall canine population (19%), with a considerably higher prevalence (43%) in the autochthonous breed of Canarian Warren hound. The prevalence in the feline population (33%) is higher than that of the canine population, and the existence of specific D. immitis antibodies in the inhabitants of the island of Gran Canaria (12%) is confirmed. In both cats and humans, the prevalence, according to the different climate areas on the island, is related to the prevalence of D. immitis in dogs in the same area, which shows the key epidemiological role played by the canine host with regard to the transmission to other hosts of D. immitis.


Subject(s)
Cat Diseases/epidemiology , Dirofilaria immitis/immunology , Dirofilariasis/epidemiology , Dog Diseases/epidemiology , Zoonoses/epidemiology , Adult , Age Factors , Animals , Antibodies, Helminth/blood , Cat Diseases/blood , Cat Diseases/parasitology , Cat Diseases/transmission , Cats , Climate , Dirofilariasis/blood , Dirofilariasis/parasitology , Dirofilariasis/transmission , Dog Diseases/blood , Dog Diseases/parasitology , Dog Diseases/transmission , Dogs , Female , Geography , Humans , Male , Middle Aged , Seroepidemiologic Studies , Spain/epidemiology , Young Adult , Zoonoses/parasitology , Zoonoses/transmission
7.
Vet Parasitol ; 176(4): 313-6, 2011 Mar 22.
Article in English | MEDLINE | ID: mdl-21310535

ABSTRACT

Cardiopulmonary biomarkers are biological parameters that can be objectively measured and quantified as indicators of pathogenic processes (heartworm disease) or as indicators of response to therapeutic intervention. To determine levels of cardiopulmonary biomarkers in canine dirofilariasis, measurements of cardiac troponin T, cardiac troponin I, myoglobin, and D-dimer concentrations were performed for dogs with and without evidence of adult heartworm infection. The results showed that levels of cardiac troponin T were undetectable in all dogs studied while levels of cardiac troponin I were higher in dogs infected with Dirofilaria immitis. In healthy dogs, levels of myoglobin and D-dimer were below detection limits of the instrument and were significantly higher in heartworm-infected dogs, notably in microfilaremic dogs. The results suggest the possibility of using troponin I and myoglobin as markers for cardiac damage and the D-dimer as a supportive tool for a diagnosis of pulmonary thromboembolism in dogs with cardiopulmonary dirofilariasis.


Subject(s)
Biomarkers/blood , Dirofilaria immitis/pathogenicity , Dirofilariasis/pathology , Dog Diseases/pathology , Myocardium/pathology , Pulmonary Embolism/diagnosis , Animals , Case-Control Studies , Dirofilariasis/diagnosis , Dirofilariasis/parasitology , Dog Diseases/diagnosis , Dog Diseases/parasitology , Dogs , Female , Fibrin Fibrinogen Degradation Products/analysis , Male , Myoglobin/blood , Troponin I/blood , Troponin T/blood
8.
J Dairy Sci ; 94(1): 410-5, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21183051

ABSTRACT

To investigate the use of sodium dodecyl sulfate (SDS) as a biocide on goat colostrum, 2 experiments were performed. In the first, 20 goat colostrum samples were divided into 3 aliquots. A different treatment was performed on each aliquot: pasteurization (56°C, 30 min) or addition of SDS to a final concentration of either 0.1 or 1% (36°C, 10 min). Immunoglobulin G and colony-forming units were evaluated before and after treatment. Both pasteurization and treatment with 1% SDS significantly reduced the colony-forming units in colostrum. Treatment with 0.1% SDS was not effective at reducing the colony-forming units in colostrum. The IgG concentration of pasteurized colostrum was significantly lower than that of untreated colostrum, whereas treatment with 1% SDS did not affect the colostrum IgG concentration. In the second experiment, the effects of SDS colostrum treatment on immune passive transfer were evaluated. Forty goat kids were fed either refrigerated colostrum or colostrum treated with 1% SDS twice daily for 2 d. Blood samples were obtained at birth and every day for 5 d. IgG, IgM, and IgA were measured in blood serum to monitor the passive immune transfer process. Creatinine, glucose, total cholesterol, blood urea nitrogen, bilirubin, and aspartate transaminase were also monitored to evaluate the health of kids. No differences in serum IgG, IgM, IgA, creatinine, glucose, total cholesterol, blood urea nitrogen, bilirubin, or aspartate transaminase levels were observed between groups. Our findings indicate that SDS is an efficient colostrum biocide that, unlike pasteurization, does not affect immune passive transfer or goat kid health.


Subject(s)
Animals, Newborn/immunology , Anti-Bacterial Agents/pharmacology , Colostrum/drug effects , Goats/immunology , Immunity, Maternally-Acquired/drug effects , Sodium Dodecyl Sulfate/pharmacology , Animals , Bacteria/drug effects , Colostrum/immunology , Colostrum/microbiology , Female , Goats/blood , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Pregnancy
9.
Vet Parasitol ; 173(1-2): 165-8, 2010 Oct 11.
Article in English | MEDLINE | ID: mdl-20605072

ABSTRACT

Canine cardiopulmonary dirofilariosis caused by Dirofilaria immitis is endemic on the island of Gran Canaria. Epidemiological surveys have shown differences in the prevalence of heartworm disease in the canine population of the island between 1989 and 1998. The aim of the present study was to follow-up the prevalence of D. immitis in both the canine population as a whole and the Canarian Warren Hound population on Gran Canaria between 2000 and 2008. Prevalences observed were always significantly higher in the Canarian Warren Hound population than in the entire canine population. A significant decrease in the prevalence (from 30.19% to 19.36%) in the whole population was observed. In the Canarian Warren Hound the prevalence decreased slightly between 2000 and 2007 (40.42-34.65%), rising in 2008 to values higher than those observed at the beginning of follow-up study (41.6%). These changes are mainly attributed to lack of preventive chemotherapy and the prolonged exposure of the Canarian Warren Hound to mosquito bites. As this breed can act as a natural reservoir in Gran Canaria, regular surveillance is needed to detect changes in the epidemiological picture on this island.


Subject(s)
Dirofilariasis/epidemiology , Dog Diseases/epidemiology , Animals , Dirofilariasis/blood , Dog Diseases/blood , Dogs , Seroepidemiologic Studies , Spain/epidemiology , Time Factors
10.
J Anim Physiol Anim Nutr (Berl) ; 92(3): 324-5, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18477313

ABSTRACT

The objective of this study was to assess the relationship between canine body condition and metabolic parameters like serum lipids, blood glucose and alanine aminotransferase (ALT) concentrations. We selected 127 dogs (42 males and 85 females) that were taken to our veterinary medicine service during routine visits. The mean age was 6.67 +/- 5.24 years. Body condition (BC) was measured by Laflamme scale and dogs were considered as obese when BC score was over 6. The following variables were collected: total cholesterol, high-density lipoprotein cholesterol, triglycerides, basal glucose and ALT. 66.1% of the dog cohort were obese. Total cholesterol and triglycerides were found to be higher (p < 0.05) in obese dogs with respect to normal weight dogs. In conclusion, obesity in dogs is associated with higher serum lipid levels.


Subject(s)
Body Composition/physiology , Dog Diseases/blood , Lipids/blood , Obesity/veterinary , Animal Feed , Animal Nutritional Physiological Phenomena , Animals , Body Constitution/physiology , Cholesterol/blood , Cholesterol, HDL/blood , Cholesterol, LDL/blood , Cross-Sectional Studies , Dog Diseases/epidemiology , Dogs , Female , Lipid Metabolism , Male , Obesity/blood , Obesity/epidemiology , Triglycerides/blood
11.
Parasitol Res ; 100(1): 103-5, 2006 Dec.
Article in English | MEDLINE | ID: mdl-16821033

ABSTRACT

Blood samples from 823 dogs were tested for circulating Dirofilaria immitis antigen during a 1-year period (May 2002 to May 2003) on Tenerife Island, Canary Islands, Spain. Seroprevalence of heartworm infection was 21%. Heartworm infection was similar in males and females and was more common in dogs aged >6 years. Distribution of infection in varying climatic zones was not statistically different.


Subject(s)
Dirofilaria/isolation & purification , Dirofilariasis/epidemiology , Dog Diseases/parasitology , Seroepidemiologic Studies , Animals , Atlantic Islands/epidemiology , Climate , Dog Diseases/epidemiology , Dogs , Incidence , Spain
12.
Vet Parasitol ; 130(1-2): 163-8, 2005 Jun 10.
Article in English | MEDLINE | ID: mdl-15893083

ABSTRACT

Trypanosoma evansi was diagnosed for the first time in the Canary Islands (Spain) in 1998 in a dromedary camel. Seroprevalences of 4.8% up to 9% have been observed using different diagnostic methods. Affected animals have been treated but the dissemination of the disease is unknown. This article presents an outbreak of abortions and high neonatal mortality attributable to T. evansi infection in camels as well as the clinical assessment of the affected animals. The patients were diagnosed by routine checking (three pregnant animals), after abortion (five dams), or after delivered premature or weak calves (eight dams). At clinical examination, 2 out of 16 affected animals showed moderate signs of chronic form, particularly hyporexia and intolerance to exercise. The aborted fetuses were aged 6-8 months of gestation, approximately. The main laboratorial findings were regenerative anemia (haemolytic anemia), lymphocytic and monocytic leukocytosis, hyperproteinemia, hyperglobulinemia, hypoglycaemia, serum urea increased and serum iron decreased. Treatment using trypanocidal drug (Cymelarsan) resulted highly effective. Massive treatment would be recommended in the entire camel population in the Canary Islands (less than 2000 animals), as therapeutic or preventive measure, in order to control and to achieve an eventual eradication of the disease.


Subject(s)
Abortion, Veterinary/parasitology , Camelus/parasitology , Disease Outbreaks/veterinary , Trypanosoma/growth & development , Trypanosomiasis/veterinary , Aborted Fetus/parasitology , Abortion, Veterinary/epidemiology , Animals , Animals, Newborn , Arsenicals/therapeutic use , Atlantic Islands/epidemiology , Female , Parasitemia/drug therapy , Parasitemia/epidemiology , Parasitemia/parasitology , Parasitemia/veterinary , Pregnancy , Trypanocidal Agents/therapeutic use , Trypanosomiasis/drug therapy , Trypanosomiasis/epidemiology , Trypanosomiasis/parasitology
13.
Vet Parasitol ; 90(1-2): 155-9, 2000 Jun 10.
Article in English | MEDLINE | ID: mdl-10828522

ABSTRACT

Trypanosomosis due to Trypanosoma evansi (surra) is a major enzootic disease of the dromedary camel. Thus, the purpose of the present study was to assess seroprevalence and infection rates in the Canary Islands using antibody(-card agglutination test-CATT/T. evansi) and parasite detection tests (micro-Haematocrit Centrifugation technique, Giemsa stained blood smears, microscopic examination of lymph node aspirates and mouse inoculation). PCV was also determined. 745 dromedary camels (483 females and 262 males) were examined. Trypanosomes were detected in seven animals. 36 animals yielded CATT positive results while 709 animals were negative. All parasitologically positive animals were also CATT positive. Results showed a good correlation between CATT positive and low PCV and a higher seroprevalence in older animals. Trypanocidal drugs have not been registered in Spain and, consequently, if vigilance is not exercised the prevalence could be increased in the future.


Subject(s)
Camelus , Protozoan Infections, Animal/epidemiology , Trypanosomiasis/veterinary , Agglutination Tests/veterinary , Animals , Atlantic Islands/epidemiology , Female , Hematocrit , Male , Mice , Sensitivity and Specificity , Seroepidemiologic Studies , Trypanosoma , Trypanosomiasis/epidemiology
15.
Prev Vet Med ; 47(1-2): 53-9, 1999 Oct 19.
Article in English | MEDLINE | ID: mdl-11018734

ABSTRACT

After the description in Grand Canary Island of a case of dromedary trypanosomosis caused by Trypanosoma evansi in 1998, an indirect enzyme immunoassay for the detection of specific anti-T. evansi IgG (Ab-ELISA) was used to assess the seroprevalence of this disease on the Canary Islands. Seroprevalence was 9.0% in the four studied islands (Gran Canaria, Tenerife, Lanzarote and Fuerteventura), varying from 10.0 to 7.5% by island (not significantly different). Prevalence using Ab-ELISA was higher than that observed when a parasitological method (microscopic observation of blood smears) was used (1.3%).


Subject(s)
Antibodies, Protozoan/blood , Camelus/parasitology , Enzyme-Linked Immunosorbent Assay/veterinary , Trypanosoma/isolation & purification , Trypanosomiasis/veterinary , Animals , Seroepidemiologic Studies , Spain/epidemiology , Trypanosoma/immunology , Trypanosomiasis/epidemiology
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