Immunization of cattle with a Rhipicephalus microplus chitinase peptide containing predicted B-cell epitopes reduces tick biological fitness.

Rhipicephalus microplus, the cattle fever tick, is the most important ectoparasite impacting the livestock industry worldwide. Overreliance on chemical treatments for tick control has led to the emergence of acaricide-resistant ticks and environmental contamination. An immunological strategy based on vaccines offers an alternative approach to tick control. To develop novel tick vaccines, it is crucial to identify and evaluate antigens capable of generating protection in cattle. Chitinases are enzymes that degrade older chitin at the time of moulting, therefore allowing interstadial metamorphosis. In this study, 1 R. microplus chitinase was identified and its capacity to reduce fitness in ticks fed on immunized cattle was evaluated. First, the predicted amino acid sequence was determined in 4 isolates and their similarity was analysed by bioinformatics. Four peptides containing predicted B-cell epitopes were designed. The immunogenicity of each peptide was assessed by inoculating 2 cattle, 4 times at 21 days intervals, and the antibody response was verified by indirect ELISA. A challenge experiment was conducted with those peptides that were immunogenic. The chitinase gene was successfully amplified and sequenced, enabling comparison with reference strains. Notably, a 99.32% identity and 99.84% similarity were ascertained among the sequences. Furthermore, native protein recognition was demonstrated through western blot assays. Chitinase peptide 3 reduced the weight and oviposition of engorged ticks, as well as larvae viability, exhibiting a 71% efficacy. Therefore, chitinase 3 emerges as a viable vaccine candidate, holding promise for its integration into a multiantigenic vaccine against R. microplus.

Investigation of Babesia spp. and Theileria spp. in ticks from Western China and identification of a novel genotype of Babesia caballi.

Babesia spp. and Theileria spp. are tick-borne protozoan parasites with veterinary importance. In China, epidemiological and genetic investigations on many Babesia and Theileria species were still absent in many areas and many tick species. From Aug 2021 to May 2023, 645 ticks were collected from the body surface of domestic animals (camels, goats, sheep, and cattle) using tweezers in seven counties in three provinces including Xinjiang (Qitai, Mulei, Hutubi, and Shihezi counties), Chongqing (Youyang and Yunyang counties), and Qinghai (Huangzhong county). Three tick species were morphologically and molecularly identified (334 Hyalomma asiaticum from Xinjiang, 245 Rhipicephalus microplus from Chongqing, and 66 Haemaphysalis qinghaiensis from Qinghai). A total of three Babesia species and two Theileria species were detected targeting the 18S gene. The COI and cytb sequences were also recovered from Babesia strains for further identification. In R. microplus from Chongqing, Babesia bigemina, the agent of bovine babesiosis, was detected. Notably, in H. asiaticum ticks from Xinjiang, a putative novel genotype of Babesia caballi was identified (0.90%, 3/334), whose COI and cytb genes have as low as 85.82% and 90.64-90.91% nucleotide identities to currently available sequences. It is noteworthy whether the sequence differences of its cytb contribute to the drug resistance of this variant due to the involvement of cytb in the drug resistance of Babesia. In addition, Theileria orientalis and Theileria annulata were detected in R. microplus from Chongqing (12.20%, 31/245) and H. asiaticum from Xinjiang (1.50%, 5/334), respectively. These results suggest that these protozoan parasites may be circulating in domestic animals in these areas. The pathogenicity of the novel genotype of B. caballi also warrants further investigation.

Characterization of cutaneous lesions caused by natural tick infestation in cattle.

Although tick infestation is a significant health problem in livestock, there are limited studies on the dermatopathological aspects of natural tick infestation in cattle. This study aimed to describe the gross and histologic aspects of cutaneous lesions caused by tick infestation in cattle. Thirteen cases were selected based on necropsy data from a 10-year retrospective study. Predispositions were observed in beef cattle (P = .049) and the Angus breed (P = .012), and lesions occurred mainly in the fall (P = .007). Gross lesions included hypotrichosis (13/13; 100%), scales (12/13; 92%), alopecia (11/13; 85%), ulcers (7/13; 54%), crusts (7/13; 54%), and erosions (2/13; 15%). These gross lesions were mainly located in the thorax (12/13; 92%), head (11/13; 85%), abdomen (10/13; 77%), neck (9/13; 69%), limbs (9/13; 69%), and perineum (9/13; 69%). Histologically, all cases had ticks adhered to the epidermis with erosions (13/13; 100%), ulcers (11/13; 85%), orthokeratotic hyperkeratosis (13/13; 100%), irregular acanthosis (13/13; 100%), intraepidermal pustules (13/13; 100%), crusts (10/13; 77%), and ballooning degeneration (4/13; 31%). In the dermis, just below the tick insertion site, there was coagulation necrosis, fibrin deposition, and inflammatory infiltrate composed of mixed cells (neutrophils, lymphocytes, plasma cells, macrophages, and few eosinophils) (9/13; 69%), neutrophils (3/13; 23%), or eosinophils (1/13; 8%). This study reinforces the different patterns of cutaneous lesions caused by tick infestation in cattle, which should be considered as a potential cause of dermatitis in this species.

Ecological factors shaping ectoparasite communities on heteromyid rodents at Médanos de Samalayuca.

Rodent ectoparasites are vectors for important pathogens of wildlife, domestic animals, and even zoonosis. Nevertheless, distribution patterns of ectoparasites are not fully understood; habitat, season, and host species are important predictors of distribution and prevalence. Heteromyid rodents are considered important reservoirs of diseases, given the presence of different ectoparasites and pathogens in them, and they offer the opportunity to learn about the ecology of parasites. The aim of the present work was to survey ectoparasites associated with heteromyid rodents near a National Protected Area in Chihuahua Mexico, south of the USA-Mexico border, and asses the effects of ecological factors (season, vegetation type, host species, and host body condition) on parasite infestation. We sampled five different locations from January 2018 to July 2022; 845 heteromyid rodents were examined and 49 fleas and 33 ticks were collected. Ectoparasites belonged to the Siphonaptera and Ixodida orders, including three families Ixodidae (Riphicephalus sanguineus), Pulicidae (Pulex irritans), and Ctenophthalmidae (Meringins altipecten, M. dipodomys). Five species of host rodents were captured, Dipodomys merriami, D. ordii, Chaetodipus eremicus, C. hispidus, and C. intermedius, but the last two species did not present any ectoparasites. Dipodomys merriami presented the highest flea and tick prevalence followed by D. ordii. We found parasitic partnerships between heteromyids according to ecological factors. The infestation in C. eremicus was related to body condition, vegetation type, and sex; in D. merriami, it was related to vegetation type and season, while D. ordii did not present a clear pattern of infestation. Our results suggest that the infestation patterns of heteromyid rodents in desert habitats are species dependent.

Current profile of phenotypic pyrethroid resistance in Rhipicephalus microplus (Acari: Ixodidae) populations sampled from Marathwada region of Maharashtra state, India.

This study examined the pattern of resistance to widely applied synthetic pyrethroids, i.e., cypermethrin and deltamethrin, against larvae of Rhipicephalus microplus ticks sampled from Marathwada region in Maharashtra, India. The study also examined the role of α- and ß-esterases and glutathione-S-transferase (GST) in resistance development. All eight R. microplus isolates tested were resistant to deltamethrin (RL IV), having RR50 values from 6.88 to 131.26. LPT analysis exhibited the resistance level II deltamethrin resistance in Beed and Hingoli, III in Dharashiv, and IV in Sambhajinagar, Parbhani, Latur, Jalna, and Nanded isolates. The LIT analysis showed that Dharashiv field isolates had the lowest LC50 value of 229.09 ppm against cypermethrin, while Sambhajinagar field isolates had the highest at 489.78 ppm. The RR50 ranged from 1145.45 to 2448.9. Seven isolates were level I resistant to cypermethrin while the Jalna isolate was level II resistant. In larvae treated with deltamethrin and cypermethrin, the activity of α- and ß-esterase enzymes increased significantly compared to control groups. The enzyme ratios in treated larvae ranged from 0.7533 to 1.7023 for α-esterase and 0.7434 to 3.2054 for ß-esterase. The Hingoli isolate treated with cypermethrin exhibited the highest α-esterase activity (903.261), whereas Sambhajinagar isolate had the highest GST enzyme ratio (2.8224) after deltamethrin exposure. When exposed to cypermethrin, the Hingoli isolate showed the highest GST enzyme ratio, 2.0832. The present study provides the current resistance status in tick populations from Marathwada region indicating deltamethrin and cypermethrin to be ineffective for tick control. The results also suggest that SP compounds should be regulated in this region and alternative control strategies should be introduced.

The complete mitochondrial genome of cattle tick clade C reveals the genetic relationship within Rhipicephalus microplus complex.

Cattle ticks (Rhipicephalus microplus) are important economic ectoparasites causing direct and indirect damage to cattle and leading to severe economic losses in cattle husbandry. It is common knowledge that R. microplus is a species complex including five clades; however, the relationships within the R. microplus complex remain unresolved. In the present study, we assembled the complete mitochondrial genome of clade C by next-generation sequencing and proved its correctness based on long PCR amplification. It was 15,004 bp in length and consisted of 13 protein genes, 22 transfer genes, and two ribosomal genes located in the two strains. There were two copies of the repeat region (pseudo-nad1 and tRNA-Glu). Data revealed that cox1, cox2, and cox3 genes were conserved within R. microplus with small genetic differences. Ka/Ks ratios suggested that 12 protein genes (excluding nad6) may be neutral selection. The genetic and phylogenetic analyses indicated that clade C was greatly close to clade B. Findings in the current study provided more data for the identification and differentiation of the R. microplus complex and made up for the lack of information about R. microplus clade C.

Competitive displacement and acaricide resistance of two Rhipicephalus (Boophilus) species collected on commercial farms in South Africa.

Rhipicephalus (Boophilus) microplus, an invasive species to Africa, and the endemic R. (B.) decoloratus are of high economic importance in the cattle industry. Invasion of the alien species in South Africa has mostly been reported for traditional communal grazing areas where it seemed to be rapid and, in some cases, even replaced the native species. The alien species is also assumed to already be resistant to acaricides upon invasion. The presence of R. (B.) microplus on commercial farms was therefore investigated and resistance screening of both species to field concentrations of cypermethrin, amitraz, and chlorfenvinphos was determined by means of the larval immersion test. Results showed that only 3.7% (of 383) tick collections submitted were R. (B.) microplus populations. A further 1.6% (of 383) showed co-existence of the two species. Comparing the level of resistance to the acaricides between the two species indicated a mean phenotypic resistance of 66.2 and 26.5% of R. (B.) decoloratus populations to cypermethrin and amitraz, respectively. This was significantly lower for R. (B.) microplus, with 23.0 and 4.1% of its populations resistant to cypermethrin and amitraz, respectively. Closed commercial farming areas seemed to have a preventative advantage for the invasion of R. (B.) microplus and displacement of R. (B.) decoloratus by R. (B.) microplus. Regular monitoring of these two species may be of high importance to prevent unnecessary financial losses due to insufficient control and increased awareness of the threat of Asiatic babesiosis vectored by R. (B.) microplus.

Metabolic rate does not scale with body size or activity in some tick species.

Respiration in ticks is highly efficient and exceptionally low. Ticks can survive years between bloodmeals by having low activity and respiration to conserve energetic resources. Our objective was to compare metabolic (VCO2) and activity rates across 6 tick species. We predicted that VCO2 would be different among species and scale linearly with activity and body mass. Activity and CO2 production were measured for 32 h in 6 tick species: Dermacentor andersoni, D. variabilis, Haemaphysalis longicornis, Rhipicephalus appendiculatus, R. microplus, and R. sanguineus. Individual ticks were measured for 30 min three times to ensure breathing occurred. Absolute and mass-specific VCO2, total activity, body mass, and ventilation patterns were compared among species. As expected, ticks did not always breathe during the 30-minute measurements, especially R. sanguineus. Ventilation patterns differed among species with R. microplus having primarily cyclic patterns and R. appendiculatus having discontinuous gas exchange. VCO2 did not scale with body mass in most species. Haemaphysalis longicornis and R. sanguineus had the lowest VCO2; however, H. longicornis was the second most active species. Life history, including questing behavior and range expansion, could be contributing to differences between species. For instance, H. longicornis had exceptionally low metabolic rates despite above average activity levels, suggesting an energetic advantage which may underlie recently documented range expansions in North America. Our results demonstrate how ticks utilize energetic resources to maximize longevity. Future research describing questing behavior and distribution modeling may help explain differences in metabolic rates and activity and impacts on life history traits.

Beauveria bassiana native strains affect the reproductive index of Rhipicephalus microplus ticks.

Rhipicephalus microplus poses a significant economic threat due to its role in transmitting Babesia bigemina, B. bovis and Anaplasma marginale. Chemical control methods, commonly employed, encounter challenges like resistance, high costs, and environmental concerns. Emerging as an alternative, entomopathogenic fungi, particularly Beauveria bassiana, present a promising avenue for biological control. Molecular identification using the internal transcribed spacer (ITS1-5.8-ITS4) region ensures accurate species identification. This study investigated two B. bassiana strains, assessing their molecular characterization, impact on R. microplus mortality, and reproductive effects on adult females. The Reproductive Aptitude Index (RAI) is employed to evaluate tick egg viability post-treatment, providing insights into the potential of these fungi for tick control. Results indicate the BbLn2021-1 strain causes 96% mortality, and BbSf2021-1 induces 100% mortality. The commercial strain exhibited 28% mortality, while the control treatment showed 12%. Statistical analysis reveals a significant difference between treatments (p < 0.01). The Reproductive Efficiency Index (REI) underscores BbSf2021-1is superiority, yielding lower egg weights than other treatments. Regarding the RAI, BbLn2021-1 and BbSf2021-1 show no significant differences but differ significantly from the commercial and control (p < 0.01). These findings suggest that strains isolated and characterized from the natural environment could have potential applications in field trials, serving as a biocontrol alternative for R. microplus ticks.

Molecular and immunological studies on Theileria equi and its vector in Egypt.

Equine piroplasmosis is not fully understood regarding pathogenicity, prophylaxis, host immune response expression, and specific vectors. Accurately identifying the parasite vector is crucial for developing an effective control plan for a particular infection. This study focused on morphologically identifying two Hyalomma species (H. anatolicum and H. marginatum) and one Rhipicephalus annulatus (R. annulatus) at the species level. The identification process was followed by phylogenetic analysis using the neighbor-joining method based on the cytochrome oxidase subunit 1 (COXI) gene as a specific vector for Theileria equi (T. equi) in horses. T. equi was diagnosed morphologically and molecularly from infected blood samples and crushed tick species using conventional PCR. Subsequently, phylogenetic analysis based on the amplification of the 18 S rRNA gene was conducted. The obtained sequence data were evaluated and registered in GenBank under accession numbers OR064161, OR067911, OR187727, and OR068139, representing the three tick species and the isolated T. equi, respectively. The study demonstrated that T. equi infection leads to immune system suppression by significantly increasing the levels of oxidative stress markers (CAT, GPx, MDA, and SOD) (P ≤ 0.0001), with this elevation being directly proportional to parasitemia levels in infected blood cells. Furthermore, a correlation was observed between parasitemia levels and the expression of immune response infection genes (IFN-gamma, TGF-ß1, and IL-1ß cytokines) in infected horses compared to non-infected equine. Common macroscopic symptoms indicating T. equi infection in horses include intermittent fever, enlarged lymph nodes (LN), and tick infestation.