Heat treatment of serum samples from stray dogs naturally exposed to Dirofilaria immitis and Dirofilaria repens in Romania

Ciucă, Lavinia; Genchi, Marco; Kramer, Laura; Mangia, Carlo; Miron, Liviu; Prete, Luisa Del; Maurelli, Maria Paola; Cringoli, Giuseppe; Rinaldi, Laura

doi:10.1016/j.vetpar.2016.05.032

Cited by 30 publications

(32 citation statements)

References 26 publications

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“…Antigen testing was performed every six months according to manufacturer’s instructions, and when the result was negative, 450 μl of the sample was heated to 104 °C for 10 min, and the resulting coagulum centrifuged [12]. The supernatant was removed and tested according to the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

Heat pretreatment of canine samples to evaluate efficacy of imidacloprid + moxidectin and doxycycline in heartworm treatment

Bendas

Mendes-de-Almeida

Simson³

et al. 2017

Parasites Vectors

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BackgroundConsidering the recent information on the increase of Dirofilaria immitis antigen detection by rapid assays in canine blood samples after heat treatment, the proposal that immune complexes block D. immitis antigen detection and that macrocyclic lactone + doxycycline (alternative protocol) might lead to increased production of those immune complexes, resulting in the erroneous diagnosis of adult worm elimination, and that there is no recommended adulticide marketed in Brazil, a study was performed to evaluate the interference of moxidectin + doxycycline (moxi-doxy) on diagnostic procedures when heartworm positive dogs are treated with this alternative protocol. Twenty-two naturally infected pet dogs were treated monthly with topical 10% imidacloprid + 2.5% moxidectin and with oral doxycycline (10 mg/kg BID/30 days) (moxi-doxy). All the dogs had their microfilaremia level determined prior to the first day of treatment, and were tested every 6 months for microfilariae (Mf) detection prior to heating, and for antigen detection prior to and after heating, the sample.ResultsThe results indicate that the treatment protocol can eliminate adult heartworms as early as 6 months after the first dose, especially in low microfilaremic dogs (< 300 Mf/ml). In this study, all dogs were free of heartworm antigen after 18–24 months of treatment. In a comparison of pre-heated samples and non-heated samples, sample pre-heating increased antigen detection sensitivity, and non-heated samples tended to be antigen-negative earlier than the pre-heated samples, especially when dogs had low microfilaremia levels. These discrepancies were not present in a subsequent sample of the same dog 6 months later.ConclusionsTwo negative antigen test results 6 months apart can be recommended as the criterion to consider when a dog has been cleared of infection. The initial microfilaremia level of a dog can be used to estimate the necessary time frame to end the treatment period.

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Section: Methodsmentioning

confidence: 99%

Heat pretreatment of canine samples to evaluate efficacy of imidacloprid + moxidectin and doxycycline in heartworm treatment

Bendas

Mendes-de-Almeida

Simson³

et al. 2017

Parasites Vectors

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“…Furthermore, antibody-complexes due to e.g., chemotherapy (Drake et al 2015) or other reasons leading to falsenegative results in current antigen-diagnosis were present in 7 % of samples in a study from the United States of America (Velasquez et al 2014). This phenomenon may be reverted by pre-treatment of sera with heat , Ciucă et al 2016. The aim of this study was the development of an ELISA to detect antibodies against Dirofilaria spp.…”

Section: Introductionmentioning

confidence: 99%

Specific Antibody Detection in Dogs with Filarial Infections

et al. 2017

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Dirofilariosis is a mosquito-transmitted disease of wild and domestic carnivores. Etiological diagnosis on canine Dirofilaria-infections is generally either based on the morphological or molecular characterization of microfilariae (L1), or in case of D. immi tis-infection on the detection of circulating antigens shed by mature female worms. However, these tests do not detect infections during the long prepatent period of 182 -238 days. We hereby present a monoclonal antibody based sandwich-ELISA used for on-plate purification of somatic antigen of adult D. immitis stages for the detection of antibodies against D. immitis and D. repens in dog sera. Sensitivity of the assay for D. immitis patent infections was calculated to be 93.8 % (95 % CI: 79.2 -99.2 %), and for patent D. repens-infections 100 % (95 % CI: 81.9 -100 %). Specificity was determined to be 98.6 % (95 % CI: 92.2 -100 %) with sera of 69 dogs from a non-endemic area. Cross-reactions against other nematodes such as Acantho cheilonema and Dipetalonema spp. (50 % and 66.7 %, respectively), Crenosoma vulpis (16.7 %), Capillaria aerophila (14.3 %) and naturally, but not experimentally infected dogs with Angiostrongylus vasorum (14.3 %) were present. However, in all positive dogs a residence in a filarial endemic area cannot be excluded. No positive reactions could be shown in experimentally infected dogs with Toxocara canis, Ancylostoma caninum and Trichuris vulpis. Dogs experimentally infected with D. repens showed seroconversion between 24 -80 days post inoculation (dpi), far earlier than beginning of patency (189 -259 dpi). Accordingly, the presented ELISA could be a supplementary or alternative tool for Original Article S82EndoparasitEs the diagnosis of Dirofilaria-infected dogs in low or non-endemic areas to document the contact rate and infection pressure.

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“…Heartworms are nematodes belonging to the Dirofilaria genus. Both D. immitis and D. repens species usually affect dogs, but D. immitis is the most important in veterinary medicine causing Canine Heartworm Disease (CHD) (CIUCA et al, 2016;SMOUT et al, 2016). It is a serious and potentially fatal disease in many regions of the world (MCCALL et al, 2008;BOWMAN;ATKINS, 2009;BORTHAKUR et al, 2016;MITERPAKOVA et al, 2016;SMOUT et al, 2016), including Brazil (OGAWA et al, 2013;LABARTHE et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

“…and Culex spp. Moreover, favorable climatic conditions as well as traffic between harmless and endemic regions also contribute to the increased number of cases (LABARTHE et al, 1997;DA SILVA;LANGONI, 2009;CIUCA et al, 2016). Intrinsic factors, ie.…”

Section: Introductionmentioning

confidence: 99%

“…The diagnosis includes direct observation of circulating microfilariae and immunodiagnostic tests (BATISTA et al, 2008;CIUCA et al, 2016). Recently, molecular diagnosis, ie polymerase chain reaction (PCR), has been shown to provide important information mainly in non-endemic areas, in which immunodiagnostic tests may fail to detect the infected animals (TARELLO, 2001;RISHNIW et al, 2006;TAYLOR et al, 2010;CIUCA et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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Canine Heartworm Disease in a Brazilian Non-Endemic Area

Monobe

Silva²,

Melanchauski³

et al. 2016

ArqVet

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Canine Heartworm Disease (CHD) is a mosquito-borne disease caused by Dirofilaria immitis. In this study, two mature adult-senior dogs from a non-endemic area to CHD presented clinical signs suggestive to the disease. The first one presented skin lesions, loss of appetite, weakness, pale mucosa membrane, and hyperthermia, whereas the second one presented severe ascites, anorexia and exercise intolerance, lateral decumbency, and marked heart murmurs. Both presented tachypnea, thrombocytopenia, leukocytosis, and microfilaremia. Multiplex-PCR (COI gene) resulted positive to D. immitis research in both cases, confirmed by sequencing, with 98% homology to D. immitis (Gen Bank accession n.AJ537512-1). In addition, both animals have never had any prophylactic treatment to CHD, and no reports about traveling to coastal areas. This study reported two unusual cases of D. immitis infection in non-endemic area from Brazil.

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Heat treatment of serum samples from stray dogs naturally exposed to Dirofilaria immitis and Dirofilaria repens in Romania

Cited by 30 publications

References 26 publications

Heat pretreatment of canine samples to evaluate efficacy of imidacloprid + moxidectin and doxycycline in heartworm treatment

Heat pretreatment of canine samples to evaluate efficacy of imidacloprid + moxidectin and doxycycline in heartworm treatment

Specific Antibody Detection in Dogs with Filarial Infections

Canine Heartworm Disease in a Brazilian Non-Endemic Area

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