Detection of winter tick, Dermacentor albipictus, infestations using near infrared reflectance spectroscopy of bovine feces.

The objective of this study was to determine whether artificial infestations of D. albipictus could be detected in cattle using near infrared reflectance spectroscopy of bovine feces (fNIRS) and if detection capability was sensitive to size of tick infestation and phase of on-host stage-specific tick development. Fecal samples were collected daily from six non-infested then later tick-infested Bos taurus yearling heifers who each served as their own control. Cattle with D. albipictus infestations arising from as few as 1000 larvae were identified by fecal chemistry changes using fNIRS technology. In two separate trials, three animal pairs were infested with one of three treatment levels (low: ∼ 1000, medium: ∼ 4000, and high: ∼ 8000) of D. albipictus larvae in a repeated measures experimental design. Trial 1 consisted of tick naïve cattle while Trial 2 consisted of prior tick exposed cattle. Date of drop and daily sum of engorged female ticks were tabulated to characterize each infestation. Cluster, common factor, principal component and MANOVA analyses were used to define and assess fecal spectra changes associated with experimental stages of infestation. Cluster analyses found significant differences in fecal samples for heifer pairs in each treatment level group (low, medium, and high) in Trial 1 and then in Trial 2 from two pre-infestation control periods (outside and inside), three stages of tick development (larval feeding, nymphal feeding, adult feeding), and post-tick recovery periods. Five shifts in fecal chemistry of non-infested and tick-infested periods were identified by six clusters of NIRS fecal spectra measured between 576 and 1126 nm. The PCA's resulted in 97.56% and 97.77% for Trials 1 and 2 respectively of the total variation in the 1050 frequencies being explained by the first three principal components (P1, P2, P3). Results from the MANOVA and the Wilk's Lambda test for both trials showed highly significant evidence (p-values < 0.0001) of a difference in the means of the three principal components across the six Stages. There was significant evidence in Trial 1 (p-values = 0.0067) and Trial 2 (p-values < 0.0001) of a difference between the means of the three principal components across the three levels of tick infestation. These significant pair-wise comparisons reflect developmental phases of tick attachment and blood-feeding that define periods of increasing, peak and declining stress identified in five fecal chemistry shifts defined by six fecal spectral clusters.

Raman-based identification of tick species (Ixodidae) by spectroscopic analysis of their feces.

Ticks are blood-feeding parasites that vector a large number of pathogens of medical and veterinary importance. There are strong connections between tick and pathogen species. Timely detection of certain tick species on cattle can cease the spread of numerous devastating diseases such as Bovine babiesiosis and anaplasmosis. Detection of ticks is currently performed by slow and laborious scout-based inspection of cattle. In this study, we investigated the possibility of identification of tick species (Ixodidae) based on spectroscopic signatures of their feces. We collected Raman spectra from individual grains of feces of seven different species of ticks. Our results show that Raman spectroscopy (RS) allows for highly accurate (above 90%) differentiation between tick species. Furthermore, RS can be used to predict the tick developmental stage and differentiate between nymphs, meta-nymphs and adult ticks. We have also demonstrated that diagnostics of tick species present on cattle can be achieved using a hand-held Raman spectrometer. These findings show that RS can be used for non-invasive, non-destructive and confirmatory on-site analysis of tick species present on cattle.

Bovine fecal chemistry changes with progression of Southern Cattle Tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) infestation.

Surveillance for cattle fever ticks is an essential activity in the U.S. Cattle Fever Tick Eradication Program which prevents reestablishment of these tick vectors of the pathogens causing bovine babesiosis. Other methods of detecting tick infested cattle could augment current physical inspection of restrained cattle by program inspectors. The objective of this study was to determine whether a single infestation of ∼5000 Rhipicephalus (Boophilus) microplus larvae induced changes in fecal chemistry that were detectable using near-infrared reflectance spectroscopy (NIRS). Fecal samples were collected daily from 6 tick-infested and 6 non-infested Bos taurus yearling heifers. Each infested animal received ticks from one of 6 different strains of laboratory colonies of R. microplus. Date of drop and daily sum of engorged female ticks were tabulated to characterize each infestation. Cluster, common factor, principal component and MANOVA analyses were used to define and assess fecal spectra changes associated with experimental stages of infestation. Cluster analyses found no significant differences in fecal samples from each of the 6 infested heifers. Two shifts in fecal chemistry of infested animals were identified by three clusters of NIRS fecal spectra. The first cluster was comprised of samples from pre-infestation to 9 days after infestation, a period inclusive of larval tick attachment and feeding. The second cluster was comprised of samples from day 10-22 corresponding to the period of nymphal feeding, adult feeding, and early drop of engorged females. A third cluster was comprised of samples from days 23-46 corresponding to the period of engorged female drop and declining tick numbers. A Tukey-Kramer multiple comparison procedure identified significant differences in fecal spectra between five experimental stages of R. microplus infestation for principal component 1 including pre-infestation to nymphal feeding, pre-infestation to adult feeding, larval feeding to adult feeding, nymphal feeding to adult feeding and nymphal feeding to engorged female drop; for principal component 2 including pre-infestation to nymphal feeding, pre-infestation to adult feeding, and pre-infestation to engorged female drop; and for principal component 3 including pre-infestation to drop, and adult feeding to drop. These significant pair-wise comparisons reflect developmental phases of tick attachment and blood-feeding that define periods of increasing, peak and declining stress identified in two fecal chemistry shifts defined by three fecal spectra clusters. Among non-infested animals, two shifts in fecal chemistry were also detected by three fecal-spectra clusters that occurred in synchrony with those of their tick-infested counterparts. There were no significant differences in principal components or MANOVA analyses between infested and non-infested animals and the pattern of significant pair-wise Tukey-Kramer multiple comparisons for non-infested animals were similar to those of infested animals. This unintended confounding effect is attributed to the manner in which all 12 animals were preconditioned as a group, then isolated in randomly assigned blind stalls in a common barn facility for the study, creating the basis for physiological stress resonance among non-infested animals.