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1.
Exp Parasitol ; 206: 107771, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31585116

RESUMO

A PCR targeting mitochondrial cytochrome oxidase subunit III (cox3) for molecular detection of Babesia gibsoni infection in dogs has been developed in this study. Fifty blood samples from suspected clinical cases from dogs, brought to the veterinary college clinics, were examined for presence of B. gibsoni using conventional diagnosis by microscopic examination of Giemsa stained thin blood smears. In addition, species specific PCRs targeting ITS-1 region (BgITS-1 PCR) and nested PCR targeting 18S ribosomal RNA gene (Bg18SnPCR) were carried out. A 634 bp PCR fragment of B. gibsoni cox3 gene was amplified in positive samples from three geographical locations of Satara, Wai and Pune in Maharashtra state of India. From analysis of the sequence of the B. gibsoni cox3 gene, we found that the Indian isolate had 96-98% similarity to the isolate from Japan and China. Post sequencing, de-novo diagnostic primer pair for species specific amplification of 164 bp fragment of B. gibsonicox3 was designed and the PCR was standardized. The diagnostic results of de-novo Bgcox3 PCR were compared with BgITS-1 PCR and Bg18S nPCR. Thin blood smears detected 22% (11/50) samples positive for small form of Babesia species. The BgITS-1 PCR detected 25% samples (15/50) as positive and Bg18S nPCR detected 80% (40/50) B. gibsoni positive samples. The de-novo Bgcox3 PCR detected 66% (33/50) samples positive for B. gibsoni (at 95% CI). The analytical sensitivity of cox3 PCR was evaluated as 0.000003% parasitaemia or 09 parasites in 100  µl of blood. The de-novo diagnostic cox3 PCR did not cross react with control positive DNA from other haemoprotozoa and rickettsia like B. vogeli, Hepatozoon canis, Trypanosoma evansi, Ehrlichia canis and Anaplasma platys. Statistically, cox3 PCR had better diagnostic efficiency than ITS-1 PCR in terms of sensitivity (p = 0.0006). No statistically significant difference between results of cox3 PCR and 18S nPCR was observed (p = 0.1760). Kappa values estimated for each test pair showed fair to moderate agreement between the observations. Specificity of Bgcox3 PCR was 100% when compared with microscopy or BgITS-1 PCR. Sensitivity of Bgcox3 PCR was 100% when compared with that of Bg18S nPCR.


Assuntos
Babesia/isolamento & purificação , Babesiose/diagnóstico , Doenças do Cão/diagnóstico , Complexo IV da Cadeia de Transporte de Elétrons/genética , Mitocôndrias/enzimologia , Animais , Babesia/classificação , Babesia/genética , Babesiose/parasitologia , Sequência de Bases , Reações Cruzadas , DNA Espaçador Ribossômico/química , Doenças do Cão/parasitologia , Cães , Eritrócitos/parasitologia , Funções Verossimilhança , Filogenia , Reação em Cadeia da Polimerase/veterinária , Valor Preditivo dos Testes , RNA Ribossômico 18S/análise , Sensibilidade e Especificidade , Alinhamento de Sequência/veterinária
2.
Ann Parasitol ; 65(2): 183-186, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31578842

RESUMO

A 10-year-old intact female Chihuahua, 2.5 kg of weight and BCS 2 (range 1­5) was taken in for medical consultation due to the presence of four skin lesion, two in the ventral thoracic region and two in the dorsal region. The dog was receiving medication due to congestive heart failure. A cutaneous form of canine leishmaniosis was diagnosed using molecular tools from a 10-years-old Chihuahua dog living in the Caribbean region. The critical health condition of the geriatric patient may have evolved to a fatal renal failure. This report is the first of a fatal case of leishmaniosis in a dog from the endemic region in Mexico.


Assuntos
Doenças do Cão , Leishmania infantum , Leishmaniose Cutânea , Animais , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Cães , Evolução Fatal , Feminino , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/patologia , Leishmaniose Cutânea/veterinária , México , Pele/parasitologia
4.
Exp Parasitol ; 206: 107768, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31539540

RESUMO

Canine leishmaniosis due to Leishmania infantum is a widespread zoonotic disease. Although aminosidine can be an effective treatment, current therapeutic recommendations do not advocate its use, mainly due to concerns regarding the potential nephrotoxicity and ototoxicity of this drug. The aim of this randomized, blinded, controlled study was to evaluate the nephrotoxicity and ototoxicity of aminosidine-allopurinol combination and compare it with that of meglumine antimonate-allopurinol combination in non-azotemic dogs with leishmaniosis. Forty dogs with leishmaniosis were randomly assigned to be treated with either aminosidine at 15 mg/kg, subcutaneously, once daily for 28 days (group A) or with meglumine antimonate at 100 mg/kg, subcutaneously, once daily for 28 days (group B). In addition to either drug, dogs in both groups were administered allopurinol at 10 mg/kg per os twice daily for 2 months. Kidney function was evaluated through measurement of serum creatinine, urea nitrogen, inorganic phosphorus, and cystatin-c concentrations and complete urinalysis, including protein-to-creatinine ratio, at baseline and after 14, 28, and 60 days from the beginning of the treatment. At the same time points, vestibular and auditory functions were evaluated through neurological examination and brainstem auditory evoked response (BAER) recordings of wave I, wave V, inter-wave I-V latencies, and minimum hearing thresholds. None of the dogs developed clinicopathological evidence of kidney disease during the study. Serum creatinine concentration increased >0.3 mg/dl over baseline in 2 dogs in group A and in 5 dogs in group B. Parameters of kidney function were not significantly different or were improved compared to baseline and the only difference between the two groups was the lower concentration of serum creatinine in group A. None of the dogs developed peripheral vestibular syndrome or hearing impairment. At the end of the study, parameters of auditory function were not significantly different or were improved compared to baseline and there were no differences between the two groups. The results of this study show that the nephrotoxicity and ototoxicity of aminosidine, when administered to non-azotemic dogs with leishmaniosis at 15 mg/kg subcutaneously once daily for 28 days along with allopurinol, is minimal and does not differ from that of meglumine antimonate.


Assuntos
Alopurinol/efeitos adversos , Doenças do Cão/tratamento farmacológico , Audição/efeitos dos fármacos , Rim/efeitos dos fármacos , Leishmaniose Visceral/veterinária , Paromomicina/efeitos adversos , Alopurinol/administração & dosagem , Alopurinol/uso terapêutico , Animais , Cóclea/efeitos dos fármacos , Creatinina/sangue , Doenças do Cão/parasitologia , Cães , Método Duplo-Cego , Combinação de Medicamentos , Potenciais Evocados Auditivos do Tronco Encefálico , Feminino , Perda Auditiva/induzido quimicamente , Perda Auditiva/veterinária , Injeções Subcutâneas/veterinária , Leishmania infantum , Leishmaniose Visceral/tratamento farmacológico , Masculino , Antimoniato de Meglumina/administração & dosagem , Antimoniato de Meglumina/efeitos adversos , Antimoniato de Meglumina/uso terapêutico , Exame Neurológico/veterinária , Paromomicina/administração & dosagem , Paromomicina/uso terapêutico , Distribuição Aleatória , Vestíbulo do Labirinto/efeitos dos fármacos
5.
Parasitol Res ; 118(10): 3011-3017, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31418113

RESUMO

Blastocystis is a ubiquitous protozoan with a wide range of hosts. In humans, its presence has been associated with gastrointestinal disorders, although its role as a pathogen still needs to be elucidated. Until now, 17 Blastocystis subtypes (STs) have been identified, with ST1-ST4 the most commonly found in humans. Among domestic animals, the same STs reported in humans have been detected in dogs. An epidemiological survey on dog kennels was carried out to evaluate the prevalence of Blastocystis and the STs involved. Overall, 99 faecal samples were collected from the rescue shelters. Blastocystis detection was performed through conventional barcoding PCR targeting the 1800-bp SSU-rDNA, followed by sequencing and phylogenetic analysis. Blastocystis DNA was found in 21 faecal samples (21.2%), and all samples were successfully sequenced and identified as ST3 in a unique monophyletic group. The presence of Blastocystis was reported for the first time in dogs from Italy, with the identification of ST3, the subtype most commonly found in humans.


Assuntos
Infecções por Blastocystis/veterinária , Blastocystis/genética , Doenças do Cão/epidemiologia , Animais , Blastocystis/classificação , Blastocystis/isolamento & purificação , Infecções por Blastocystis/diagnóstico , Infecções por Blastocystis/epidemiologia , Infecções por Blastocystis/parasitologia , DNA de Protozoário/genética , DNA Ribossômico/genética , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Cães , Fezes/parasitologia , Feminino , Itália/epidemiologia , Masculino , Epidemiologia Molecular , Filogenia , Prevalência , Análise de Sequência de DNA
6.
Parasitol Res ; 118(10): 2979-2987, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31435764

RESUMO

Enterocytozoon bieneusi is an obligate intracellular protist-like fungi parasite that infects numerous mammal hosts including humans, raising concerns of zoonotic transmission. There is little information available on the presence and diversity of E. bieneusi genotypes in companion animals. Here, we determined the occurrence and genetic diversity of E. bieneusi in domestic dogs and cats from Northern Spain. A total of 336 genomic DNA samples extracted from canine (n = 237) and feline (n = 99) faecal specimens were retrospectively investigated. The presence of E. bieneusi was assessed by PCR of the rRNA internal transcribed spacer (ITS) gene. The parasite was detected in 3.0% (3/99) and 0.8% (2/237) of the cats and dogs examined, respectively. All three feline positive samples were from stray cats living in an urban setting, whereas the two canine samples were from owned dogs living in rural areas. Sequence analysis revealed the presence of two genotypes in dogs, BEB6 and PtEb IX, and two genotypes in cats, D and Peru11. The identification of Peru11 in a cat and BEB6 in a dog constitutes the first report of those genotypes in such hosts as well as first report in Spain. This is also the first evidence of genotype D in cats and PtEb IX in dogs in Spain. Three out of the four genotypes, BEB6, D and Peru11, have been previously reported as human pathogens and are potentially zoonotic indicating that dogs and cats need to be considered potential sources of human infection and environmental contamination.


Assuntos
Doenças do Gato/parasitologia , Doenças do Cão/parasitologia , Enterocytozoon/genética , Variação Genética , Microsporidiose/veterinária , Animais , Doenças do Gato/epidemiologia , Gatos , Doenças do Cão/epidemiologia , Cães , Enterocytozoon/isolamento & purificação , Fezes/parasitologia , Genótipo , Microsporidiose/epidemiologia , Microsporidiose/parasitologia , Estudos Retrospectivos , Espanha/epidemiologia
7.
Parasit Vectors ; 12(1): 380, 2019 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-31362754

RESUMO

BACKGROUND: Canine heartworm disease is a potentially fatal disease for which treatment is financially burdensome for many pet owners. Prevention is strongly advocated by the veterinary community along with routine testing for infection during annual wellness examinations. Despite the availability of efficacious chemoprophylaxis, recent reports have suggested that the incidence of heartworm disease in domestic dogs is increasing. RESULTS: Using data from tests for heartworm infection in the USA from January 2012 through September 2018, a Bayesian spatio-temporal binomial regression model was used to estimate the regional and local temporal trends of heartworm infection prevalence. The area with the largest increase in regional prevalence was found in the Lower Mississippi River Valley. Regional prevalence increased throughout the southeastern states and northward into Illinois and Indiana. Local (county-level) prevalence varied across the USA, with increasing prevalence occurring along most of the Atlantic coast, central United States, and western states. Clusters of decreasing prevalence were present along the Mississippi Alluvial Plain (a historically endemic area), Oklahoma and Kansas, and Florida. CONCLUSIONS: Canine heartworm infection prevalence is increasing in much of the USA, both regionally and locally, despite veterinarian recommendations on prevention and testing. Additional steps should be taken to protect dogs, cats and ferrets. Further work is needed to identify the driving factors of the locally decreasing prevalence present along the Mississippi Alluvial plain, Florida, and other areas.


Assuntos
Dirofilariose/epidemiologia , Doenças do Cão/epidemiologia , Cães/parasitologia , Animais de Estimação/parasitologia , Animais , Antígenos de Helmintos/sangue , Dirofilaria immitis , Dirofilariose/diagnóstico , Doenças do Cão/parasitologia , Feminino , Geografia , Hospitais Veterinários/estatística & dados numéricos , Incidência , Mississippi/epidemiologia , Modelos Estatísticos , Prevalência , Análise Espaço-Temporal , Estados Unidos/epidemiologia
8.
Ann Parasitol ; 65(2): 145-149, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31376344

RESUMO

The aim of this paper was to analyse the amino acid sequences of the 18S rRNA gene of Babesia canis strains and the proteomic analysis of the serum of dogs infected with three various genotypes: 18S rRNA B. canis. Material for the research was DNA B. canis obtained from dogs with babesiosis. In total, 60 DNA tested samples were divided into three groups (20 samples each). The groups were formed by DNA samples of the sequences marked as 18S RNA-A (group 1), 18S RNA-B (group 2), and 18S RNA-C (group 3). The basis for the classification of protozoa to a specific group was the location of relevant nucleotides (GA, AG, or TT) in position 150-151 of the tested nucleotide sequence 18S rRNA. Nucleotide sequences were transcribed into amino acid sequences and then analysed using DNASTAR software. From all 60 infected and ten healthy dogs (control group), the serum was taken to make proteomic tests using MALDI-TOF mass spectrometer. It was demonstrated that the mutations found in position 150 and 151 of the nucleotide sequence, result in a change of amino acid sequences. Moreover, it was also demonstrated that the disease course in dogs infected with different strains of protozoa is different. Each of the analysed strains of protozoa induced in the serum of infected animals the appearance of a protein fraction of mass 51 kDa, which may then be treated as a nonspecific disease marker used for the diagnosis of this disease but not to differentiate the protozoa strains.


Assuntos
Babesia , Babesiose , Doenças do Cão , Proteoma , RNA de Protozoário , RNA Ribossômico 18S , Sequência de Aminoácidos , Animais , Babesia/genética , Babesiose/sangue , Babesiose/parasitologia , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Cães , Proteínas de Protozoários/sangue , Proteínas de Protozoários/química , RNA de Protozoário/química , RNA Ribossômico 18S/química , RNA Ribossômico 18S/genética
9.
Ann Parasitol ; 65(2): 151-158, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31378013

RESUMO

Dogs which spend their lives in rural areas are primarily "sentinels" of animal herds. The dogs' close contact with humans and other animals obligates their caregivers to systematic deworming. The goal is to eliminate potential contamination with parasites which is mostly caused by direct contact or food and water contaminated with dogs' excretions. The aim of this study was to assess internal parasitic invasions on dogs from rural areas which spend most of their lives on farms. In total, 69 samples of fresh stools were collected for the analysis, including 26 from females and 43 from males. Coprological analysis of stool samples was accompanied with a survey where pet owners gave information on the number of visits to the veterinarian in suspicion of their dogs being infected with the parasites, the number of deworming treatments in the previous year and the dogs' behavior towards strangers. Also, the age, sex and body weight of dogs were recorded. Statistical analysis of the obtained results was performed using Statistica 12.5 with a medical set by Statsoft. In total, 29 (46,03%) out of 69 fecal samples contained developmental forms of parasites. Research has shown that animals most vulnerable to invasion were in the 6­10 years old age group, mostly males. In addition, the invasion of Toxocara canis roundworms occurred most frequently, as it was found in 12 (41.37%) animals (6 females and 6 males). The analysis of the questionnaires showed that the dog owners' knowledge of the risk of infection and parasites is unsatisfactory. This corresponded with a small number of deworming treatments. It is significant that nearly half of the owners never dewormed their dogs.


Assuntos
Doenças do Cão , Parasitos , Animais , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Fezes/parasitologia , Feminino , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Masculino , Parasitos/isolamento & purificação , População Rural/estatística & dados numéricos , Inquéritos e Questionários , Toxocara canis , Toxocaríase/epidemiologia
10.
BMC Infect Dis ; 19(1): 747, 2019 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-31455227

RESUMO

BACKGROUND: Leishmaniasis caused by different species of Leishmania affect 98 countries worldwide. Visceral Leishmaniasis (VL) is the mortal clinical presentation of the disease that causes the dead to more than 90% of the patients who suffer it. The diagnosis of VL is made by the direct observation of the parasite in bone marrow, spleen and/or liver aspirates that requires complex proceedings. Therefore, serum samples are submitted to Indirect Immunofluorescence to identify the presence of anti-Leishmania antibodies. Despite the variability in the diagnostic performance of the Immunochromatographic Tests (ICTs), there are many evidences that suggest that ICTs can be used for epidemiological screening. However, in Colombia there are not any evidence about the performance of the ICTs for VL diagnosis, both for human and canine serum samples. Therefore, this study evaluated the diagnostic performance of 4 ICTs for VL (2 ICTs in human sera and 2 ICTs in canine sera) in samples from endemic areas of Colombia. METHODS: We selected a total of 156 human serum samples (82 positive and 74 negative for VL) and 126 canine serum samples (71 positive and 54 negative) diagnosed by in house Indirect Immunofluorescence (IIF). The samples were submitted to the ICTs following the manufacturers' instructions. Statistical analysis was performed to evaluate the diagnostic performance of each ICT in comparison with the IIF. PCR for HSP70 gene and sanger sequencing was performed in samples with negative results for both ICTs. RESULTS: The sensitivity (S) of both ICTs for human samples (Ad-bio Leishmania IgG/IgM Combo Rapid Test and Kalazar Detect™) was 91.5% and specificity (E) were 93.2 and 89.2% respectively, while for the ICTs tested on canine samples (Kalazar Detect™ Rapid Test, Canine and DPP® CVL rapid test) we found S values between 82.9 and 85.7% and E values between 79.6 and 92.6%. We found L. infantum by PCR and sequencing in 2 human samples, and L. braziliensis and L. amazonensis in canine serum samples that were negative by both ICTs. CONCLUSIONS: We conclude that both tests evaluated on human samples have a similar diagnostic performance, while the Kalazar Detect™ Rapid Test, Canine showed a better diagnostic performance than the DPP® CVL rapid test evaluated on canine samples. Also, we suggest that it is necessary to design tests with antigens of the circulating strains to increase its diagnostic utility.


Assuntos
Doenças do Cão/diagnóstico , Imunoensaio/métodos , Leishmaniose Visceral/diagnóstico , Animais , Colômbia , Testes Diagnósticos de Rotina , Doenças do Cão/parasitologia , Cães , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Leishmania braziliensis/genética , Leishmania braziliensis/imunologia , Leishmania infantum/genética , Leishmania infantum/imunologia , Leishmaniose Visceral/veterinária , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade
11.
Parasit Vectors ; 12(1): 369, 2019 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-31349867

RESUMO

BACKGROUND: The efficacy of an extended-release injectable moxidectin (0.5 mg/kg) suspension (ProHeart® 12) (PH 12) in preventing the development of Dirofilaria immitis in dogs for 12 months was investigated in laboratory and field studies in the USA. METHODS: In each of two laboratory studies, 20 dogs ≥ 12 months of age were randomly allocated to receive a subcutaneous injection of saline or PH 12 on Day 0 and were then inoculated with 50 D. immitis third-stage larvae (L3) on Day 365. All dogs were necropsied ~ 5 months post-inoculation for adult worm counts. The field efficacy study included dogs ≥ 10 months of age from 19 veterinary clinics in the USA treated with either 20 monthly doses of Heartgard® Plus (HG Plus) (296 dogs) or two doses of PH 12 (297 dogs) on Days 0 and 365. Efficacy was determined on Days 365, 480 and 605 using adult HW antigen and microfilaria testing to assess adult HW infection. RESULTS: PH 12 was 100% effective in preventing HW disease in all three of these studies. In the laboratory studies, no PH 12-treated dogs had any adult HWs, whereas all control dogs in both studies had adult HWs [geometric mean, 30.2 (range, 22-37) for Study 1 and 32.6 (22-44) for Study 2]. In the field study, all dogs treated with PH 12 tested negative for adult HW infection on all test days (Days, 365, 480 and 605), whereas four dogs receiving HG Plus (positive control) tested positive for HWs during the study (three dogs on Day 365 and one dog on Day 480). All four dogs treated with HG Plus that subsequently tested positive for HWs during the field study were from the lower Mississippi River Valley region, where HW resistance to macrocyclic lactone preventives has been confirmed to occur. PH 12 was significantly better than HG Plus in preventing heartworm disease in the field study (P = 0.0367). PH 12 was well-tolerated in both laboratory and field studies. CONCLUSIONS: A single dose of ProHeart® 12 was 100% effective in preventing heartworm disease in dogs for a full year in both laboratory and field studies.


Assuntos
Dirofilaria immitis/efeitos dos fármacos , Dirofilariose/prevenção & controle , Doenças do Cão/prevenção & controle , Filaricidas/administração & dosagem , Macrolídeos/administração & dosagem , Animais , Preparações de Ação Retardada/administração & dosagem , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Cães , Feminino , Hospitais Veterinários/estatística & dados numéricos , Injeções Subcutâneas , Masculino , Distribuição Aleatória , Estados Unidos
12.
Prev Vet Med ; 169: 104696, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31311632

RESUMO

Canine babesiosis is a serious disease among tick-borne haemoprotozoan diseases, globally. The present study was envisaged for carrying out thorough investigation of the disease among working dogs of organised kennels situated in different agro-climatic zones of India as comprehensive understanding of the disease from this country was pertinently lacking. During the study period of three years (2012-2014), 330 dogs suspected for babesiosis were examined for clinicopathology by their physical examination, haematological and biochemical parameters estimation, while the detection of apicomplexan parasites was confirmed by using various diagnostic techniques i.e. by conventional microscopy, by two different Babesia specific 18S rRNA based PCR protocols (conventional/simple PCR and nested PCR assays) followed by sequencing of obtained PCR amplicons for Babsesia spp. identification. Out of 330 clinical cases screened 5.15% (17/330), 9.09% (30/330) and 15.45% (51/330) were found to be positive in microscopic examination, simple- and nested- PCR assay, respectively. Comparative statistical analyses of these diagnostic assay results revealed that significant difference exists among the three diagnostic methodologies and thus it is recommended that the nested PCR technique be relied upon as a screening molecular assay and also for epidemiological studies of the disease in this country. Phylogenetic analysis based on 18S rRNA depicted the monophyletic nature and clonal expansion among all the B. gibsoni, under study. Sequencing results of PCR amplicons revealed that B. gibsoni has predominantly established itself over B. vogeli as former was incriminated in 47 cases while latter was confirmed in only four animals. Based on the clinical severity, these 51 affected animals were classified into three main groups' of 17 animals each viz., apparently healthy-, simple or uncomplicated babesiosis- and atypical or complicated babesiosis- group. Haematological and biochemical profiling of these dogs confirmed the characteristics findings of infection by both the Babesia spp. It was observed that the infection by small form of Babesia (B. gibsoni) is posing a significant therapeutic challenge and chemosterilization by commonly prescribed anti-protozoal drugs was not achieved as clinical relapses were often observed. The clinical signs, sequence based confirmation and severity of the infection suggested that there is a positive selection of B. gibsoni (smaller form) over B. vogeli (larger form) in this country and raises serious concerns as prognosis in former is considered to be poor compared to latter. Thus, these findings have opened new paradigms for planning of pragmatic control strategies against this emerging canine health problem.


Assuntos
Babesia/genética , Babesiose/epidemiologia , Babesiose/genética , Doenças do Cão/epidemiologia , Doenças do Cão/genética , Animais , Babesia/isolamento & purificação , Babesiose/sangue , Babesiose/patologia , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Cães , Feminino , Abrigo para Animais , Índia/epidemiologia , Masculino , Epidemiologia Molecular , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 18S/genética , Análise de Sequência de DNA
13.
Artigo em Inglês | MEDLINE | ID: mdl-31300132

RESUMO

Infectious haemolytic anaemia (IHA) in dogs share similar clinical signs including fever, lethargy, icterus, paleness of mucous membranes and splenomegaly. Postmortal findings are similar and, without additional diagnostic methods, an accurate aetiological diagnosis is difficult to achieve. In order to investigate causes of lethal IHA in Croatian dogs, we performed a retrospective study on archived formalin-fixed, paraffin-embedded tissue blocks (FFPEB) from dogs that died due to haemolytic crisis, using microscopic and molecular diagnostic tools to determine the aetiological cause of disease. Molecular analysis was performed on kidney, lung, myocardium and spleen on FFPEB from all dogs. The originally stated aetiological diagnosis of B. canis or leptospirosis was confirmed in only 53% of the dogs. PCR and sequencing revealed that, in addition to the expected pathogens, B. canis and Leptospira interrogans, the presence of previously undiagnosed "new" pathogens causing anaemia including Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum. Furthermore, Theileria capreoli was detected for the first time in a dog with postmortal descriptions of lesions. Intensive extravascular hemolysis was noticeable as jaundice of the mucosa, subcutis and fat tissue, green or yellow discoloration of renal parenchyma caused by bilirubin excretion in the renal tubules and bile accumulation within the liver in 90% of the dogs. This work highlights the value of molecular diagnostics to complement traditional ante-mortem and post-mortem diagnostic protocols for the aetiological diagnosis of pathogens associated with IHA.


Assuntos
Anemia Hemolítica/veterinária , Doenças do Cão/diagnóstico , Ehrlichiose/veterinária , Doenças Transmitidas por Carrapatos/veterinária , Anaplasmataceae/genética , Anemia Hemolítica/diagnóstico , Anemia Hemolítica/mortalidade , Animais , Autopsia/veterinária , Doenças do Cão/microbiologia , Doenças do Cão/mortalidade , Doenças do Cão/parasitologia , Cães , Ehrlichiose/diagnóstico , Feminino , Hemólise , Masculino , Técnicas de Diagnóstico Molecular , Inclusão em Parafina , Estudos Retrospectivos , Theileria/genética , Doenças Transmitidas por Carrapatos/diagnóstico
14.
Exp Appl Acarol ; 78(3): 431-442, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31270640

RESUMO

Tick-borne spotted fever in Brazil is known to be caused by two agents, Rickettsia rickettsii and Rickettsia parkeri. Nothing was known about these agents in one area of the Atlantic rainforest biome of Bahia state, where during March to June 2016, 356 rural dogs and 69 horses were sampled and their sera were processed through indirect immunofluorescence assay against antigens of R. rickettsii, R. parkeri, Rickettsia amblyommatis and Rickettsia bellii. Ticks collected from these dogs and horses were molecularly tested for the presence of rickettsial DNA. Overall, 16.4% (58/356) dogs and 24.6% (17/69) horses were seroreactive to Rickettsia spp. Five tick species, Rhipicephalus sanguineus sensu lato (s.l.), Amblyomma ovale, A. sculptum, R. microplus, and A. naponense, were collected from dogs, whereas horses were infested by A. sculptum and Dermacentor nitens. A total of 242 ticks from dogs and 62 from horses were analyzed by PCR targeting rickettsiae, which were detected in only 4/27 (14.8%) A. ovale. Fragments of the rickettsial gltA and ompA genes from these four ticks were 100% identical to the Atlantic rainforest strain of R. parkeri. The presence of A. ovale on dogs was positively associated with local canine seroreactivity to R. parkeri. Our results provide evidence for the transmission of R. parkeri strain Atlantic rainforest from A. ovale to domestic dogs within the rural area of Ilhéus, similarly to other areas of the Atlantic rainforest biome of Brazil, where human cases of R. parkeri-caused spotted fever have been reported.


Assuntos
Doenças do Cão/epidemiologia , Doenças dos Cavalos/epidemiologia , Ixodidae/microbiologia , Infecções por Rickettsia/veterinária , Rickettsia/isolamento & purificação , Infestações por Carrapato/veterinária , Animais , Brasil/epidemiologia , DNA Bacteriano/análise , Doenças do Cão/microbiologia , Doenças do Cão/parasitologia , Cães , Feminino , Doenças dos Cavalos/microbiologia , Doenças dos Cavalos/parasitologia , Cavalos , Ixodidae/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Larva/microbiologia , Masculino , Ninfa/crescimento & desenvolvimento , Ninfa/microbiologia , Prevalência , Rhipicephalus/crescimento & desenvolvimento , Rhipicephalus/microbiologia , Infecções por Rickettsia/epidemiologia , Infecções por Rickettsia/microbiologia , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/parasitologia
15.
Res Vet Sci ; 125: 205-211, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31260840

RESUMO

The factors associated with Leishmania spp. infection in dogs are still poorly understood. This study aimed to identify such factors among domestic dogs from the Brazilian municipality of Rondonópolis, which recently emerged as an endemic area with intense transmission of human visceral leishmaniasis (VL). It was a household-based cross-sectional study conducted between 2016 and 2017. Interviews were conducted with 405 dog owners considering their socioeconomic characteristics, and environmental aspects of the household and its adjacent areas. In addition, 600 dogs were evaluated regarding physical characteristics, behavior, and care provided by the owner. Seropositive animals were those that showed reactivity in a rapid immunochromatographic test and enzyme-linked immunosorbent assay, which are currently recommended for canine visceral leishmaniasis (CVL) diagnosis in Brazil. A multivariable logistic regression analysis was employed to determine the associated factors. Low social class [OR = 2.0; 95%CI = (1.2-3.2)], presence of acerola tree (Malpighia emarginata) in the yard [OR = 2.2; 95%CI = (1.2-4.1)], presence of more than one dog in the household [OR = 2.0; 95%CI = (1.3-3.3)], horse and/or cattle breeding [OR = 12.0; 95%CI = (1.9-73.6)], existence of houses with yard adjacent to the home [OR = 4.0; 95%CI = (1.3-12.2)], presence of apparent signs consistent with CVL [OR = 10.6; 95%CI = (5.8-19.4)], dog staying mostly in the yard during the day [(OR = 4.8; 95%CI = (1.1-21.4)], and lack of cleaning of the dog's shelter [(OR = 1.9; 95%CI = (1.1-3.2)] were identified as the factors associated with Leishmania spp. infection. These results support the importance of socioeconomic and environmental aspects in the occurrence of Leishmania spp. infection. In addition, they may be useful in guiding control strategies in areas where zoonotic VL is endemic.


Assuntos
Doenças do Cão/parasitologia , Leishmania/isolamento & purificação , Leishmaniose Visceral/veterinária , Animais , Brasil/epidemiologia , Estudos Transversais , Doenças do Cão/diagnóstico , Doenças do Cão/epidemiologia , Cães , Doenças Endêmicas , Ensaio de Imunoadsorção Enzimática/veterinária , Humanos , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , Propriedade , Fatores Socioeconômicos
16.
Res Vet Sci ; 125: 218-226, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31280121

RESUMO

Leishmaniosis due to Leishmania infantum is a complex infection that can affect both humans and dogs, and present a wide range of clinical signs and clinicopathological abnormalities. The conventional treatment of this disease is challenging due to the fact that complete parasitological cure commonly does not occur. Furthermore, treatment of the disease with the conventionally used drugs has several shortcomings. These include the need for long-term treatment, side effects and the formation of drug resistance. Moreover, it is important to highlight that the host immune responses play a crucial role in the outcome of this infection. For this reason, the use of immunotherapy in clinical leishmaniosis to improve the result of treatment with the conventional anti-leishmanial drugs by enhancing the immune response is imperative. The aim of this review is to provide a comparative overview of the wide range of immunotherapeutical approaches and strategies for the treatment of L. infantum infection in animals focusing on dogs.


Assuntos
Doenças do Cão/parasitologia , Imunoterapia/veterinária , Leishmaniose/veterinária , Animais , Doenças do Cão/terapia , Cães , Leishmania , Leishmaniose/terapia
17.
Vet Parasitol ; 271: 22-30, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31303199

RESUMO

Babesia rossi causes the most severe clinical disease in dogs of all the babesia parasites. We included 320 naturally-infected dogs that presented for care at the Onderstepoort Veterinary Academic Hospital between 2006 and 2016. All dogs had mono-infections confirmed by multiplex PCR. The data allowed more accurate clinical classification of the disease and identified parameters that were associated with disease severity and death. Odds ratios for dying were significant (P < 0.05) for increased band neutrophil count, collapse at presentation; presence of cerebral signs; hypoglycaemia; hyperlactatemia; high urea, high creatinine; hyperbilirubinaemia; hypercortisolaemia; and hypothyroxinaemia. Joint component analysis confirmed that the variables with significant odds ratios grouped together with death. Yet, multivariate logistic regression was unable to identify a group of significant independent predictors of death. Receiver Operator Characteristic curves indicated that low total thyroid hormone, high bilirubin, high serum urea and high cortisol concentrations were the variables with the highest sensitivity and specificity for death. These data provide both the clinician and researcher with a set of easily-measured laboratory and clinical assessments to classify cases into those that are uncomplicated and those that are complicated. The disease is complex and multisystemic and probably involves mechanisms more proximal in the pathogenesis than those that have been evaluated.


Assuntos
Babesiose/patologia , Babesiose/parasitologia , Doenças do Cão/patologia , Doenças do Cão/parasitologia , Animais , Babesia , Babesiose/epidemiologia , Babesiose/mortalidade , Doenças do Cão/epidemiologia , Doenças do Cão/mortalidade , Cães , Razão de Chances , África do Sul/epidemiologia
18.
Vet Parasitol ; 271: 38-44, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31303201

RESUMO

A nucleoside triphosphate diphosphohydrolase-1 (NTPDase 1) was identified on the surface, flagellum and kinetoplast from L. infantum promastigotes by immunocytochemistry and confocal laser scanning microscopy, using immune sera that recognized specifically the B domain of NTPDase 1 and produced against synthetic peptides (LbB1LJ and LbB2LJ) derived from this domain. The polyclonal antibodies had effective antileishmanial effect, reducing significantly in vitro promastigotes growth (21-25%), an antiproliferative effect also demonstrated by immune sera produced against recombinant r-pot B domain, and two other synthetic peptides (potB1LJ and potB2LJ). In addition, using these biomolecules in ELISA technique, IgG1 and IgG2 subclasses reactivities of either healthy dogs or infected by L. infantum and classified clinically as asymptomatic, oligosymptomatic and symptomatic were tested. Analysis of distinct IgG1 and IgG2 seropositivities patterns suggested antibody subclasses binding epitopes along B domain for protection against infection, indicating this domain as a new tool for prophylactic and immunotherapeutic investigations.


Assuntos
Anticorpos Antiprotozoários/imunologia , Doenças do Cão/imunologia , Imunoglobulina G/imunologia , Leishmania infantum/enzimologia , Leishmania infantum/imunologia , Leishmaniose Visceral/veterinária , Nucleosídeo-Trifosfatase/imunologia , Animais , Anticorpos Antiprotozoários/metabolismo , Doenças do Cão/parasitologia , Cães , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Domínios Proteicos/imunologia
19.
Vet Parasitol ; 271: 87-97, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31303211

RESUMO

The natural history of canine visceral leishmaniasis (CVL) has been well described, particularly with respect to the parasite load in different tissues and immunopathological changes according to the progression of clinical forms. The biomarkers evaluated in these studies provide support for the improvement of the tools used in developing vaccines against CVL. Thus, we describe the major studies using the dog model that supplies the rationale for including different biomarkers (tissue parasitism, histopathology, hematological changes, leucocytes immunophenotyping, cytokines patterns, and in vitroco-culture systems using purified T-cells subsets and macrophages infected with L. infantum) for immunogenicity and protection evaluations in phases I and II applied to pre-clinical and clinical vaccine trials against CVL. The search for biomarkers related to resistance or susceptibility has revealed a mixed cytokine profile with a prominent proinflammatory immune response as relevant for Leishmania replication at low levels as observed in asymptomatic dogs (highlighted by high levels of IFN-γ and TNF-α and decreased levels in IL-4, TGF-ß and IL-10). Furthermore, increased levels in CD4+ and CD8+ T-cell subsets, presenting intracytoplasmic proinflammatory cytokine balance, have been associated with a resistance profile against CVL. In contrast, a polyclonal B-cell expansion towards plasma cell differentiation contributes to high antibody production, which is the hallmark of symptomatic dogs associated with high susceptibility in CVL. Finally, the different studies used to analyze biomarkers have been incorporated into vaccine immunogenicity and protection evaluations. Those biomarkers identified as resistance or susceptibility markers in CVL have been used to evaluate the vaccine performance against L. infantum in a kennel trial conducted before the field trial in an area known to be endemic for visceral leishmaniasis. This rationale has been a guiding force in the testing and selection of the best vaccine candidates against CVL and provides a way for the veterinary industry to register commercial immunobiological products.


Assuntos
Biomarcadores/sangue , Doenças do Cão/sangue , Leishmaniose Visceral/veterinária , Animais , Biomarcadores/análise , Suscetibilidade a Doenças/metabolismo , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Leishmaniose Visceral/sangue , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Vacinas Protozoárias/imunologia
20.
Exp Appl Acarol ; 78(2): 231-246, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31152319

RESUMO

Ticks are haematophagous arthropods that exert direct and indirect effects on their hosts. Their global importance as reservoirs and vectors of diseases of veterinary and public health importance is well recognized. However, the level of understanding of their role in disease epidemiology varies from one country to the other based on available data. Information on ticks infesting dogs across Nigeria and the public health significance is scarce. Therefore, this study aimed to provide information on ixodid ticks infesting dogs in Nigeria. Ticks were collected from 608 owned dogs presented to veterinary clinics and hospitals in 10 out of 36 states of Nigeria over a 14-month period and identified using taxonomic descriptions and morphological keys. In all, 1196 ticks belonging to three genera were identified. Rhipicephalus (including the subgenus Boophilus) ticks were collected from dogs from all the states surveyed and accounted for 95.2% of the ticks collected, followed by Haemaphysalis (3.7%) and Amblyomma species (1.2%). The brown dog tick, Rhipicephalus sanguineus sensu lato was the only tick identified in all the climatic zones of Nigeria. There is a statistically significant association between tick infection rate and rainy season, female animals, local and cross breed against exotic animals, total lack of control practice by dog owners, frequency of the control and with traditional methods of tick control but not the age of the dogs. The epidemiological and public health implications of these findings were discussed.


Assuntos
Distribuição Animal , Doenças do Cão/epidemiologia , Ixodidae/fisiologia , Saúde Pública , Infestações por Carrapato/veterinária , Animais , Doenças do Cão/parasitologia , Cães , Feminino , Ixodidae/crescimento & desenvolvimento , Larva/fisiologia , Masculino , Nigéria/epidemiologia , Ninfa/fisiologia , Prevalência , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/parasitologia
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