No abstract

The purpose of this study was to describe an unreported entomopathogenic fungus that naturally infects the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). Engorged female ticks, showed symptoms of fungal infection after controlled tick infestation of cattle. Infected ticks developed a distinctive dark colour, a pale mould grew over the cuticle and the ticks eventually died covered with fungal conidiophores. The responsible fungus was isolated and cultured on mycological medium and submitted to microscopic morphology, biochemical phenotyping and 18S rRNA ribotyping analyses, which identified it as aflatoxin-producing Aspergillus flavus. Spores from the cultured fungus were experimentally sprayed over healthy engorged female ticks, obtaining an 80% prevalence of experimental infection of healthy ticks and their egg masses, the larval progeny after incubation under laboratory conditions was also infected. These results demonstrate that A. flavus is the causative agent of the natural fungal disease of the cattle tick R. microplus described here.

The goal of the present study was to assess the gene expression of xenobiotic metabolizing enzymes (XMEs) Cytochrome P-450 (CYP) and carboxylesterase (CE) related to detoxification of synthetic pyrethroids, plus acetylcholinesterase (AChE), in field isolates of acaricide-resistant Rhipicephalus microplus. The XMEs expression levels were assessed by mRNA measurement using quantitative reverse transcription PCR. The XME expression levels of field-isolated acaricide-resistant ticks were compared against acaricide-susceptible reference ticks used in this study as a gene expression baseline and represented as relative expression units (REU). Field isolates were subjected to toxicological bioassays and determined resistant to all the Pyr acaricides (Pyr), whereas most of them were found susceptible to organophosphorous acaricides (OP), with the exception of three isolates, which exhibited moderate resistance to Diazinon. Significantly higher levels of CYP were detected in pyrethroid-resistance ticks when compared to Su ticks (P<0.01). A linear regression analysis showed that pyrethroid acaricide resistance levels of R. microplus were proportional to the CYP expression levels (correlation coefficient (R):0.85; P<0.05). Analysis on CE expression levels showed only one isolate resistant to Pyr and OP with a statistically significant increase (P<0.01). AChE expression levels showed statistically significant (P<0.01) subexpression in all tick isolates when compared to the susceptible reference. Our results suggest that pyrethroid acaricide resistance in the cattle tick may be diagnosed by measuring the CYP expression levels using quantitative PCR.

Tick-borne bacterial pathogens (TBBPs) show a worldwide distribution and represent a great impact on public health. The brown dog tick (Rhipicephalus sanguineus) is a vector of several pathogens that affect dogs and sometimes humans as well. In addition, TBBPs represent a diagnostic challenge and imply financial resources and medical treatment for long periods of time. In the present study, R. sanguineus s. l. was identified as the main tick species naturally parasitizing dogs that inhabit. Juárez City, Chihuahua, in the Paso del Norte region, Mexico–US Border, representing 99.8% of the cases. Additionally, an end-point PCR was performed to search for whether pathogens in R. sanguineus s. l. can transmit in DNA extracted from ticks and dog blood samples. This is the first molecular detection of Rickettsia rickettsi infecting domestic dogs in Mexico; however, other pathogens were also identified, such as Ehrlichia canis and Anaplasma platys in both ticks and dog blood samples, while Anaplasma phagocytophilum was identified only in dog blood samples. Moreover, co-detection in tick pools and co-infection in the analyzed dog blood samples could be found. Similarly, this research showed that dogs were found mostly parasitized by adult female ticks, increasing the possibility of transmission of E. canis.

Different laboratory cultures of the acarine tick Rhipicephalus (Boophilus) microplus (Canestrini, 1888) (Ixodida: Ixodidae) were infested by small Megaselia scalaris (Loew, 1866) (Diptera: Phoridae) flies. Larvae of this species exhibited opportunistic parasitism predominantly on engorged female ticks, causing severe damage to their cuticle through which the flies were able to reach R. microplus internal organs, on which they fed until developing into pupae in the tick's remains. The flies were kept by continuous propagation on fresh ticks over six generations during which the same parasitoid behaviour was observed. Here we report on an ixodid tick laboratory culture used for rearing M. scalaris.