New records of Rhipicephalus linnaei infected by Rickettsia massiliae from Central Mexico.

BACKGROUND: On the American continent, Rhipicephalus sanguineus s.l. comprises two species: Rhipicephalus linnaei and R. sanguineus s.s. Each species has been identified as a potential vector of at least one of five species of pathogenic bacteria of the genus Rickettsia. In particular, Rickettsia massiliae is one of three species with the greatest importance in public health at the continental level. In Mexico, this species is reported exclusively in the Nearctic states of Baja California and Chihuahua. AIM: For this reason, the aim of this work was to provide new records of R. massiliae for the centre of the country derived from active acarological surveillance. MATERIALS AND METHODS: During the period of February-October 2019, 29 dogs from six municipalities in the state of Morelos were sampled. Hosts were visually inspected, and ticks were recovered and identified morphologically and molecularly by amplification of the 16S rDNA gene. Subsequently, five genes from members of the genus Rickettsia were amplified and sequenced. RESULTS: A total of 229 (117♀, 98♂ and 14 N) ticks identified as R. linnaei were recovered, two of which were positive for R. massiliae strains related to those recovered from Argentina and the United States. CONCLUSIONS: This work provides the second record of R. massiliae infecting R. linnaei in Mexico and the Americas, increasing the geographic distribution of this Rickettsia species in the Neotropical region, and providing information on the possible role of R. linnaei as a potential vector of this microorganism.

Mutational landscape and intra-host diversity of human papillomavirus type 16 long control region and E6 variants in cervical samples.

Human papillomavirus genotype 16 (HPV16) is the most frequent high-risk HPV (HR-HPV) identified in cervical precursor lesions and cervical cancer (CC) worldwide. The oncogenic potential of HPV16 is partly dependent on the lineage involved in the infection and the presence of clinically relevant mutations. In this report, we present the distribution of HR-HPV and the mutational profile and intra-host variability of HPV16 lineages, based on analysis of the long control region (LCR) and the E6 gene in samples with normal cytology (n = 39), squamous intraepithelial lesions (n = 25), and CC (n = 39). HR-HPV genotyping was performed using multiplex real-time PCR. HPV16 lineage assignments and mutation frequencies were determined by conventional PCR and Sanger DNA sequencing, and intra-patient viral populations were analyzed using next-generation sequencing (NGS). The most frequent HR-HPV type was HPV16, followed by HPV31 and HPV18. The frequency of HPV16 sublineages was A1/A2 > D2 > D3 and B1. Moreover, the most frequent mutations, both in samples from this study and in the available sequences from Mexican isolates in the GenBank database were LCR-G7518A, which is involved in carcinogenesis, and E6-T350G (producing L83V), associated with persistence of infection. Otherwise, deep sequencing revealed high conservation of viral lineages and mutations, independently of the stages studied. In conclusion, the high frequency and stability of these molecular markers, as well as the circulating viral lineages, could be related to the incidence of CC associated with HPV16. Hence, they deserve a broader analysis to determine the risk of specific populations for progression of the disease.