Resistance profile and molecular characterization of pyrethroid resistance in a Rhipicephalus microplus strain from Colombia.

Intensive use of chemical acaricides for the control of cattle ticks (Rhipicephalus microplus) has led to the development of multiple acaricide resistance in Colombia. The present study aimed to characterize, using toxicological bioassays and molecular biology techniques, the resistance profile of a tick strain isolated from the Arauca state, Northeast Colombia. Commercial acaricides were used in adult immersion tests to determine its in vitro efficacies. Deltamethrin showed very low activity (4-7.3%), a mixture of cypermethrin and chlorpyrifos had intermediate efficacy (64-75.2%), and ethion presented the highest activity (88.5-100%). A colony (Arauquita strain) was established and larval immersion tests confirmed high resistance level to deltamethrin (241-fold) and susceptibility to ivermectin. A quantitative polymerase chain reaction-high resolution melt technique was used to identify single nucleotide polymorphisms (SNPs) in the para-sodium channel gene. All of the genotyped individuals were mutant, presenting one (n = 7), two (n = 7) or three (n = 9) SNPs previously associated with pyrethroid resistance. Sequencing revealed a novel mutation (F712L), that was found for the first time in R. microplus ticks from South America. This is the first description of mutations associated with pyrethroid resistance in R. microplus from Colombia. The acaricide resistance pattern found in the Arauquita strain is similar to other parts of Colombia.

Bartonella infection in urban and rural dogs from the tropics: Brazil, Colombia, Sri Lanka and Vietnam.

Dogs can be infected by a wide range of Bartonella spp., but limited studies have been conducted in tropical urban and rural dog populations. We aimed to determine Bartonella antibody prevalence in 455 domestic dogs from four tropical countries and detect Bartonella DNA in a subset of these dogs. Bartonella antibodies were detected in 38 (8·3%) dogs, including 26 (10·1%) from Colombia, nine (7·6%) from Brazil, three (5·1%) from Sri Lanka and none from Vietnam. DNA extraction was performed for 26 (63%) of the 41 seropositive and 10 seronegative dogs. Four seropositive dogs were PCR positive, including two Colombian dogs, infected with B. rochalimae and B. vinsonii subsp. berkhoffii, and two Sri Lankan dogs harbouring sequences identical to strain HMD described in dogs from Italy and Greece. This is the first detection of Bartonella infection in dogs from Colombia and Sri Lanka and identification of Bartonella strain HMD from Asia.

Serological survey of Leishmania infantum and Trypanosoma cruzi in dogs from urban areas of Brazil and Colombia.

Leishmania infantum and Trypanosoma cruzi are zoonotic parasites that are endemic throughout many parts of Latin America. Infected dogs play an important role in transmission of both parasites to humans. A serological survey of Leishmania and Trypanosoma infection was conducted on 365 dogs from São Paulo, Brazil and Bogatá, Colombia, South America. Serum samples were examined by the indirect immunofluorescent antibody test (IFAT). Anti-Leishmania IgG antibodies were detected in 5 of 107 from Brazil (4.7%) and in 4 of 258 dogs (1.6%) from Colombia. Titers ranged from 1:25 to 1:100. Anti-T. cruzi antibodies were not detected in any of the dogs from either Brazil or Colombia. The results show a low prevalence of anti-Leishmania antibodies and no antibodies against T. cruzi in these canine populations. Our study suggests that dogs play a limited role in the spread of L. infantum and T. cruzi in these urban areas of Brazil and Colombia.

Prevalence of Toxoplasma gondii in dogs from Colombia, South America and genetic characterization of T. gondii isolates.

The prevalence of Toxoplasma gondii in 309 unwanted dogs from Bogotá, Colombia, South America was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT) and found in 52 (16.8%) of 309 dogs with titers of 1:20 in 20, 1:40 in six, 1:80 in 17, 1:160 in three, 1:320 in three, 1:1280 or higher in three. Some organs obtained after necropsy of dogs (hearts, tongues and brains, either separately or pooled) were used in bioassays carried out in mice (37 samples, of which 20 were assayed with separate organs and 17 were assayed with pooled organs), cats (pooled organs from six) and pooled organs of two dogs both in mice and cat. Mice receiving dog tissues were examined for T. gondii infection. Feces of cats that received dog tissues were examined for oocyst shedding. In total, T. gondii was isolated from tissues of 20 dogs (16 by bioassays in mice, 3 by bioassay in cats and 1 by bioassay in mice and cat). All infected mice from 7 of 17 isolates bioassayed in this host died of toxoplasmosis during primary infection. Only 10 of the 20 dogs whose tissues were bioassayed separately induced infections in mice. Interestingly, dog organs varied in their capacity to induce T. gondii infection in mice, hearts and tongues producing more positive results than the brain. The 20 T. gondii isolates obtained from seropositive dogs were PCR-RFLP genotyped using polymorphisms at 10 nuclear markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, a new SAG2 and an apicoplast marker Apico. Ten genotypes were revealed. These genotypes are different from the three predominant Types I, II and III lineages that are widely spread in North America and Europe. A new allele denoted u-3 at PK1 locus was identified in three isolates. This result supports previous findings that T. gondii population is highly diverse in Colombia.

Serological survey for antibodies to Encephalitozoon cuniculi in ownerless dogs from urban areas of Brazil and Colombia.

There are 3 strains of Encephalitozoon cuniculi that occur in mammals. Strain III is associated with clinical disease in dogs, although some can be asymptomatic carriers and excrete spores in their urine. Several cases of human E. cuniculi infection caused by strain III have been observed in immunocompromised patients, indicating that E. cuniculi should be considered a zoonotic agent. Encephalitozoon cuniculi can cause fatal disease in maternally-infected or young dogs. Clinical signs in these animals included blindness, encephalitis, retarded growth rate, and nephritis. Encephalitozoon cuniculi has also been associated with primary renal failure in adult dogs. The present study used the direct agglutination test (DAT, cut-off 1:50) and the indirect fluorescent antibody test (IFAT, cut-off 1:10) to examine the prevalence of antibodies to E. cuniculi in dogs from Brazil and Colombia. Using the DAG, 31 (27.4%) of 113 dogs from Brazil and 47 (18.5%) of 254 dogs from Colombia were seropositive. Nine (14.3%) of 63 dogs from Brazil and 18 (35.3%) of the 51 dogs from Colombia were seropositive by indirect immunofluorescent antibody test. These results indicate that dogs from Brazil and Colombia are exposed to E. cuniculi.