The Application of PCR and Reverse Line Blot Hybridization to Detect Arthropod‐borne Hemopathogens of Dogs and Cats in Trinidad

Abstract

Arthropod-borne diseases are important causes of morbidity and mortality of companion animals in Trinidad. As clinical signs are vague, more sensitive methods to diagnose these diseases based on the polymerase chain reaction (PCR) followed by reverse line blot hybridization (RLB) of amplified products are being developed. An RLB of 14 oligonucleotide probes coupled with polymerase chain-amplified regions of 16S rRNA or 18S rRNA genes of hemoparasites from cats and dogs detected Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, feline mycoplasmas ("Candidatus Mycoplasma haemominutum,"Mycoplasma haemofelis), and some unknown species within the Babesia/Theileria group and the Anaplasma/Ehrlichia tribe. Amplified products were obtained from blood samples collected from 348 dogs and 15 cats. Overall, hemopathogen DNA was detected in 92 (26.4%) dogs and six (40.0%) cats. E. canis (49, 14.1%) and feline mycoplasma (5, 33.3%) DNA were most frequently identified in dogs and cats, respectively. B. canis vogeli (1, 6.7%) and E. canis (1, 6.7%) were also detected in cats. Mixed infections of Anaplasma/Ehrlichia sp. and Babesia sp. were present in five (1.4%) dogs, while mixed infections of the feline mycoplasmas were present in two (13.3%) cats, one of which was also positive for E. canis. Pyrexia was significantly associated with a positive RLB result in dogs (P= 0.00, chi(2), 1 df). This is the first reported application of macro-arraying techniques to detect arthropod-borne hemopathogens of companion animals in the Americas and the first detection of DNA of B. canis vogeli and E. canis in cats in Trinidad.

Tick-borne agents in domesticated and stray cats from the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil

“…Feline piroplasmosis is associated with anorexia, weight loss, gastrointestinal alterations, anemia, thrombocytopenia, leukocytosis or leukopenia, and jaundice (Criado-Fornelio, 2012a; Vilhena et al, 2013). Our findings also emphasize the occurrence of B. vogeli, a canid piroplasmid, in cats in Brazil, corroborating with previous studies conducted in Brazil others regions of the world (Georges et al, 2008;Simking et al, 2010;Vilhena et al, 2013). Additionally, the present work highlights the diversity of piroplasmids circulating in cats in Brazil and the need for molecular confirmation of piroplasmas found in erythrocytes of cats' blood smears.…”

“…Recently, B. vogeli was detected in blood samples from stray domestic cats in a Brazilian zoo . Also, B. vogeli DNA has already been detected in cat's blood samples from Trinidad (Georges et al, 2008), Thailand (Simking et al, 2010) and Portugal (Vilhena et al, 2013). André et al (2011) reported the detection of Babesia spp.…”

Investigação molecular de Ehrlichia spp. e Anaplasma platys em felinos domésticos: alterações clínicas, hematológicas e bioquímicas

“…No entanto, contrariando os resultados deste estudo, alguns autores obtiveram sucesso na ampliϐicação de DNA de Ehrlichia spp. em gatos domésticos (Beauϐils et al 2002, Yin-Chiachun et al 2003, Oliveira 2008, Tabar et al 2008, Georges et al 2008) e em felinos selvagens onde André (2008) foi o pioneiro a detectar a presença de E. canis em 11 felinos selvagens mantidos em zoológicos de São Paulo e Brasília. Breitschwerdt et al (2002) Após testar a nested PCR com primers baseados na seqüência do gene 16SrRNA da Ehrlichia spp.…”

Molecular detection of Anaplasma platys in a naturally-infected cat in Brazil

“…However, E. canis DNA has been detected in blood of domestic cats in North America (2) and recently in Trinidad by reverse line blot hybridization (13). Moreover, experimental infection of cats by Neorickettsia risticii (formerly Ehrlichia risticii) has been described (8).…”

“…Blood-borne pathogens in domestic cats have been attracting the attention of researchers (11,27,13,28). With the use of molecular tools, a large number of pathogens previously recognized only as parasites of dogs have also been detected in domestic cats (2,25).…”