Artificial infestation of white-tailed deer with ticks (Acari: Ixodidae) to study tick-host interactions.

White-tailed deer (Odocoileus virginianus) are a main host for the adult life stages of tick species of medical and veterinary importance. Since white-tailed deer play a vital role in tick ecology, research has been conducted to understand this tick-host relationship. To date, research involving captive white-tailed deer and artificial infestation of these animals with ticks has focused on host suitability, the role of white-tailed deer in tick-borne diseases, and anti-tick vaccine research. The methodology reported for these studies was at times not descriptive and inconsistent regarding how and what region of the white-tailed deer was infested with ticks. Here, we propose a standardized method to artificially infest captive white-tailed deer with ticks for research purposes. The protocol describes a method proven effective to experimentally infest captive white-tailed deer with blacklegged ticks (Ixodes scapularis) to study tick-host interactions. The methods can be reliably transferred for experimental infestation of white-tailed deer by other multi-host and one-host tick species.

The genome of the stable fly, Stomoxys calcitrans, reveals potential mechanisms underlying reproduction, host interactions, and novel targets for pest control.

BACKGROUND: The stable fly, Stomoxys calcitrans, is a major blood-feeding pest of livestock that has near worldwide distribution, causing an annual cost of over $2 billion for control and product loss in the USA alone. Control of these flies has been limited to increased sanitary management practices and insecticide application for suppressing larval stages. Few genetic and molecular resources are available to help in developing novel methods for controlling stable flies. RESULTS: This study examines stable fly biology by utilizing a combination of high-quality genome sequencing and RNA-Seq analyses targeting multiple developmental stages and tissues. In conjunction, 1600 genes were manually curated to characterize genetic features related to stable fly reproduction, vector host interactions, host-microbe dynamics, and putative targets for control. Most notable was characterization of genes associated with reproduction and identification of expanded gene families with functional associations to vision, chemosensation, immunity, and metabolic detoxification pathways. CONCLUSIONS: The combined sequencing, assembly, and curation of the male stable fly genome followed by RNA-Seq and downstream analyses provide insights necessary to understand the biology of this important pest. These resources and new data will provide the groundwork for expanding the tools available to control stable fly infestations. The close relationship of Stomoxys to other blood-feeding (horn flies and Glossina) and non-blood-feeding flies (house flies, medflies, Drosophila) will facilitate understanding of the evolutionary processes associated with development of blood feeding among the Cyclorrhapha.

Feeding and reproductive parameters of adult female Ixodes scapularis (Acari: Ixodidae) and Amblyomma americanum parasitizing white-tailed deer (Odocoileus virginianus).

White-tailed deer Odocoileus virginianus (Zimmermann) (Artiodactyla: Cervidae) are the main host for adult Ixodes scapularis Say (Acari: Ixodidae) (blacklegged tick) and all stages of Amblyomma americanum Linnaeus (Acari: Ixodidae) (lone star tick). However, literature describing the feeding and reproductive parameters of these tick species when feeding on this host is limited. We experimentally infested white-tailed deer with adult pairs of either I. scapularis or A. americanum to improve our understanding of these tick-host relationships. Our study used tick-naïve white-tailed deer and restricted host grooming throughout the infestation. For I. scapularis, the days to repletion (mean ±â€…SE, 6.04 ±â€…0.07), engorgement weight of replete females (0.20 ±â€…0.0032 g), duration of oviposition (32 ±â€…0.45 d), egg mass weight (0.10 ±â€…0.0027 g), and number of eggs laid per tick (1,803.00 ±â€…49.00) were recorded. Data from A. americanum were also recorded, including days to repletion (11.00 ±â€…0.063), engorgement weight of replete females (0.63 ±â€…0.025 g), duration of oviposition (37.00 ±â€…1.30 d), egg mass weight (0.34 ±â€…0.017 g), and number of eggs laid per tick (5,873.00 ±â€…291.00). These biological parameter data could be used as variables in models (e.g., LYMESIM 2.0) to determine how white-tailed deer influence I. scapularis and A. americanum populations in nature, and to evaluate the protective efficacy of tick-antigen-based antitick vaccines.

Nilgai antelope display no signs of infection upon experimental challenge with a virulent Babesia bovis strain.

BACKGROUND: Bovine babesiosis is caused by infection with the protozoal parasite Babesia bovis, which is transmitted by Rhipicephalus (Boophilus) spp. It can cause mortality rates up to 90% in immunologically naive Bos taurus cattle. In south Texas, R. (B.) microplus is known to infest nilgai antelope (Boselaphus tragocamelus); however, their susceptibility to infection with B. bovis and their role in the transmission of the parasite remain unknown. In this study, we challenged nilgai antelope with B. bovis and evaluated their susceptibility to infection. METHODS: Nilgai were needle inoculated with ≈108 B. bovis-parasitized erythrocytes (merozoites) or a homogenate of B. bovis-infected larval ticks (sporozoite) delivered intravenously. Bos taurus beef calves were inoculated in parallel, as this strain of B. bovis is lethal to cattle. Temperature and hematocrit were monitored daily over the course of each study, and whole blood was collected for molecular [polymerase chain reaction (PCR)] and serological [indirect enzyme-linked immunosorbent assay (ELISA)] diagnostic evaluation. Histological sections of nilgai cerebral tissue were examined for evidence of infection. Recipient bovine calves were sub-inoculated with blood from nilgai challenged with either stage of the parasite, and they were monitored for clinical signs of infection and evaluated by a PCR diagnostic assay. Red blood cells (RBCs) from prechallenged nilgai and B. taurus beef cattle were cultured with an in vitro B. bovis merozoite culture to examine colonization of the RBCs by the parasite. RESULTS: Nilgai did not display clinical signs of infection upon inoculation with either the merozoite or sporozoite stage of B. bovis. All nilgai were PCR-negative for the parasite, and they did not develop antibodies to B. bovis. No evidence of infection was detected in histological sections of nilgai tissues, and in vitro culture analysis indicated that the nilgai RBCs were not colonized by B. bovis merozoites. Cattle subinoculated with blood from challenged nilgai did not display clinical signs of infection, and they were PCR-negative up to 45 days after transfer. CONCLUSIONS: Nilgai do not appear to be susceptible to infection with a strain of B. bovis that is lethal to cattle. Tick control on these alternative hosts remains a critical priority, especially given their potential to disseminate ticks over long distances.

Developmental stage and level of submersion in water impact the viability of lone star and winter tick eggs during flooding.

Female ticks deposit large egg clusters that range in size from hundreds to thousands. These clusters are restricted to a deposition site, usually under leaf litter and other debris. These sites can be exposed to periodic flooding, where the cluster of tick eggs can float to the surface or remain underneath organic debris entirely underwater. Here, we examined the viability of egg clusters from winter ticks, Dermacentor albipictus , and lone star ticks, Amblyomma americanum , when partially submerged or fully submerged in water in relation to the developmental stages of the eggs. In general, egg clusters that were older and partially submerged had a higher viability than fully submerged, young eggs in water. A. americanum was much more resistant to water exposure between the two species. These studies highlight that egg clusters for specific tick species can remain viable when exposed to water for at least two weeks, where eggs float on the surface. These studies also suggest that water-based distribution of egg clusters could occur for some species, and flooding will differentially impact tick egg survival based on the specific developmental stage of exposure and species.

Discovery of microRNAs of the stable fly (Diptera: Muscidae) by High-throughput sequencing.

The stable fly, Stomoxys calcitrans (L.), is a serious ectoparasite affecting animal production and health of both animals and humans. Stable fly control relies largely on chemical insecticides; however, the development of insecticide resistance as well as environmental considerations requires continued discovery research to develop novel control technologies. MicroRNAs (miRNAs) are a class of short noncoding RNAs that have been shown to be important regulators of gene expression across a wide variety of organisms, and may provide an innovative approach with regard to development of safer more targeted control technologies. The current study reports discovery ad initial comparative analysis of 88 presumptive miRNA sequences from the stable fly, obtained using high-throughput sequencing of small RNAs. The majority of stable fly miRNAs were 22-23 nt in length. Many miRNAs were arthropod specific, and several mature miRNA sequences showed greater sequence identity to miRNAs from other blood-feeding dipterans such as mosquitoes rather than to Drosophilids. This initial step in characterizing the stable fly microRNAome provides a basis for further analyses of life stage-specific and tissue-specific expression to elucidate their functional roles in stable fly biology.

Sequence polymorphism in acetylcholinesterase transcripts and genotyping survey of BmAchE1 in laboratory and Mexican strains of Rhipicephalus (Boophilus) microplus.

ABSTRACT Acetylcholinesterase cDNAs, BmAChE1, BmAChE2, and BmAChE3 of Rhipicephalus (Boophilus) microplus (Canestrini) were sequenced and found to exhibit significant polymorphism. A portion of the predicted amino acid substitutions in BmAChE1, BmAChE2, and BmAChE3 were found predominantly in organophosphate-resistant strains, but most did not correlate with resistant status. Multiple transcripts were observed from individual ticks, suggesting possible gene duplication or alternative splicing to produce more than two transcripts per individual. BmAChE1 transcript polymorphisms associating with organophosphate-resistant status in laboratory strains were surveyed in laboratory and Mexican strains of R. microplus by sequencing BmAChE1 genomic DNA. Quantitative real-time polymerase chain reaction was used to determine copy numbers of BmAChE1 (eight copies/haploid genome), BmAChE2 (16 copies/haploid genome), and BmAChE3 (four copies/haploid genome). Presence of at least three highly polymorphic amplified genes expressing AChE in tick synganglion suggested that ticks maintain a large and diverse assortment of AChE alleles available for rapid recombination and selection, which potentially reduces fitness costs associated with individual mutations. Elevated copy numbers for each of the BmAChEs may also explain previous failures to identify mutations resulting in insensitivity to organophosphates. It is clear that development of phenotypic resistance to organophosphates is highly complex and may be multigenic in character.

Analysis of doramectin in the serum of repeatedly treated pastured cattle used to predict the probability of cattle fever ticks (Acari: Ixodidae) feeding to repletion.

Analysis of doramectin concentration in blood serum of pastured cattle injected repeatedly (12 treatments) at two different dosage rates and 28-day intervals throughout the year was used to predict the probability that cattle fever ticks could successfully feed to repletion during the interval between any two consecutive treatments. Treatment at ~270 µg/kg indicated that serum doramectin concentration dropped below the baseline concentration estimated for tick survival (8 ppb) in 7 of the 12 treatments. However, the longest period between any two treatments during which the doramectin concentration remained below the 8 ppb baseline level for successful tick feeding was 15 days, making it virtually impossible for any ticks to reach ovipositional status prior to a subsequent treatment. At a dosage rate of ~540 µg/kg, the concentration dropped below the baseline tick survival level (8 ppb) only once, following the initial treatment, and the duration during which the concentration remained below the baseline level prior to the subsequent treatment was only 6 days. Thus, at the high dosage rate results indicated, with absolute certainty, that no ticks could successfully feed to repletion between any two consecutive treatments. Based on the data obtained in the study it was concluded that analysis of doramectin concentration in serum of treated animals would be a reliable predictor for assessing the probability that ticks could successfully develop to repletion. More importantly, results demonstrated that the trial policy, instituted by the Cattle Fever Tick Eradication Program, of repeatedly treating cattle with doramectin injections at 25-28 day intervals for eliminating cattle fever ticks would produce little or no risk of any viable ticks developing to repletion and re-infesting the field between treatment applications.

Patterns of deer ked (Diptera: Hippoboscidae) and tick (Ixodida: Ixodidae) infestation on white-tailed deer (Odocoileus virginianus) in the eastern United States.

BACKGROUND: White-tailed deer (Odocoileus virginianus) host numerous ectoparasitic species in the eastern USA, most notably various species of ticks and two species of deer keds. Several pathogens transmitted by ticks to humans and other animal hosts have also been found in deer keds. Little is known about the acquisition and potential for transmission of these pathogens by deer keds; however, tick-deer ked co-feeding transmission is one possible scenario. On-host localization of ticks and deer keds on white-tailed deer was evaluated across several geographical regions of the eastern US to define tick-deer ked spatial relationships on host deer, which may impact the vector-borne disease ecology of these ectoparasites. METHODS: Ticks and deer keds were collected from hunter-harvested white-tailed deer from six states in the eastern US. Each deer was divided into three body sections, and each section was checked for 4 person-minutes. Differences in ectoparasite counts across body sections and/or states were evaluated using a Bayesian generalized mixed model. RESULTS: A total of 168 white-tailed deer were inspected for ticks and deer keds across the study sites. Ticks (n = 1636) were collected from all surveyed states, with Ixodes scapularis (n = 1427) being the predominant species. Counts of I. scapularis from the head and front sections were greater than from the rear section. Neotropical deer keds (Lipoptena mazamae) from Alabama and Tennessee (n = 247) were more often found on the rear body section. European deer keds from Pennsylvania (all Lipoptena cervi, n = 314) were found on all body sections of deer. CONCLUSIONS: The distributions of ticks and deer keds on white-tailed deer were significantly different from each other, providing the first evidence of possible on-host niche partitioning of ticks and two geographically distinct deer ked species (L. cervi in the northeast and L. mazamae in the southeast). These differences in spatial distributions may have implications for acquisition and/or transmission of vector-borne pathogens and therefore warrant further study over a wider geographic range and longer time frame.

Identification of a mutation associated with permethrin resistance in the para-type sodium channel of the stable fly (Diptera: Muscidae).

The insect sodium channel is of particular interest for evaluating resistance to pyrethroids because it is the target molecule for this major class of neurotoxic insecticides. The stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), sodium channel coding sequence representing domains IS6 through IVS6 was isolated, and the sequence encoding domain II was compared among individuals of a laboratory strain selected for resistance to permethrin and the unselected, parental generation. A point mutation resulting in a leucine-to-histidine amino acid change was identified (Leul014His), and its location corresponded with that observed for knockdown resistance (kdr) mutations in other insects. As a result, the allele was designated kdr-his. A molecular assay was developed to assess the frequency of this mutation in genomic DNA of individual stable flies from the laboratory selections, which provided further evidence that the kdr-his allele accounts for the observed level ofpermethrin resistance in the selected strain. The assay was then used to evaluate the frequency of the mutation from five field-collected populations originating from three horse farms near Ocala, FL; one horse farm near Gainesville, FL; and one dairy farm near Hague, FL. Frequency of the kdr-his allele ranged from 0.46 to 0.78, supporting further investigation of allele prevalence throughout the stable fly season and in response to field insecticide application.

Sudden Mortality in Captive White-Tailed Deer With Atypical Infestation of Winter Tick.

In October 2020, three captive male white-tailed deer, Odocoileus virginianus [Zimmermann] (artiodactyla: Cervidae), were found dead in central Pennsylvania and a fourth was euthanized due to extreme lethargy. The deer presented with high burdens of Dermacentor albipictus (Packard) (Ixoda: Ixodidae) (winter tick). There were no other clinical symptoms and deer were in otherwise good physical condition with no observed alopecia. Winter tick epizootics have been associated with mortalities of moose, Alces alces [Linnaeus] (artiodactyla: cervidae), and more recently elk, Cervus canadensis [Erxleben] (artiodactyla: cervidae), in Pennsylvania, but have not been reported in white-tailed deer. Mild winters are favorable to winter ticks and deer producers and managers should be aware of possible infestations as a result.

Molecular screening for rickettsial bacteria and piroplasms in ixodid ticks surveyed from white-tailed deer (Odocoileus virginianus) and nilgai antelope (Boselaphus tragocamelus) in southern Texas.

A survey of ixodid ticks parasitizing white-tailed deer (Odocoileus virginianus) and nilgai antelope (Boselaphus tragocamelus) was completed during the 2018-2019 public hunt season on the Laguna Atascosa National Wildlife Refuge (Cameron County, Texas) and the East Foundation's El Sauz Ranch in nearby Willacy County (Texas). Anocenter nitens was the predominant tick species identified with 5% of these ticks collected from nilgai. All life stages were encountered in high numbers on white-tailed deer, indicating that deer may be a primary host in this region. Amblyomma maculatum and Amblyomma inornatum were identified from both hosts, while Ixodes scapularis was encountered only on white-tailed deer. This is the first published record of A. inornatum on nilgai. A subset of ticks was used in PCR assays to detect Rickettsia spp., family Anaplasmataceae, Borrelia spp., and Theileria-Babesia spp. Borrelia spp. were not detected in any of the ticks analyzed. Rickettsia parkeri was detected in three A. maculatum adult ticks from deer, Rickettsia sp. endosymbiont sequences were present in all I. scapularis ticks, and Rickettsia amblyommatis was detected in three A. inornatum adult ticks from deer. Sequence analysis of Anaplasmataceae-positive amplicons from A. nitens and A. maculatum had low percent identity to published Anaplasma spp. sequences, suggesting a unique Anaplasma sp. may be circulating in the population. Anaplasma platys was detected from A. nitens larvae and an Ehrlichia sp. Delta strain was present in A. maculatum, both of unknown pathogenicity towards deer. Theileria cervi was detected in all stages of A. nitens ticks, and positive ticks originated from 27 of 31 deer and a single nilgai sampled from throughout the survey site. The primary vector for T. cervi is absent from this region, suggesting T. cervi is possibly maintained by a different tick species.

Sex Chromosome Evolution in Muscid Flies.

Sex chromosomes and sex determining genes can evolve fast, with the sex-linked chromosomes often differing between closely related species. Population genetics theory has been developed and tested to explain the rapid evolution of sex chromosomes and sex determination. However, we do not know why the sex chromosomes are divergent in some taxa and conserved in others. Addressing this question requires comparing closely related taxa with conserved and divergent sex chromosomes to identify biological features that could explain these differences. Cytological karyotypes suggest that muscid flies (e.g., house fly) and blow flies are such a taxonomic pair. The sex chromosomes appear to differ across muscid species, whereas they are conserved across blow flies. Despite the cytological evidence, we do not know the extent to which muscid sex chromosomes are independently derived along different evolutionary lineages. To address that question, we used genomic and transcriptomic sequence data to identify young sex chromosomes in two closely related muscid species, horn fly (Haematobia irritans) and stable fly (Stomoxys calcitrans). We provide evidence that the nascent sex chromosomes of horn fly and stable fly were derived independently from each other and from the young sex chromosomes of the closely related house fly (Musca domestica). We present three different scenarios that could have given rise to the sex chromosomes of horn fly and stable fly, and we describe how the scenarios could be distinguished. Distinguishing between these scenarios in future work could identify features of muscid genomes that promote sex chromosome divergence.

Association of Salivary Cholinesterase With Arthropod Vectors of Disease.

Acetylcholinesterase (AChE) was previously reported to be present in saliva of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), with proposed potential functions to 1) reduce acetylcholine toxicity during rapid engorgement, 2) modulate host immune responses, and 3) to influence pathogen transmission and establishment in the host. Potential modulation of host immune responses might include participation in salivary-assisted transmission and establishment of pathogens in the host as has been reported for a number of arthropod vector-borne diseases. If the hypothesis that tick salivary AChE may alter host immune responses is correct, we reasoned that similar cholinesterase activities might be present in saliva of additional arthropod vectors. Here, we report the presence of AChE-like activity in the saliva of southern cattle ticks, Rhipicephalus (Boophilus) microplus; the lone star tick, Amblyomma americanum (Linnaeus); Asian tiger mosquitoes, Aedes albopictus (Skuse); sand flies, Phlebotomus papatasi (Scopoli); and biting midges, Culicoides sonorensis Wirth and Jones. Salivary AChE-like activity was not detected for horn flies Haematobia irritans (L.), stable flies Stomoxys calcitrans (L.), and house flies Musca domestica L. Salivary cholinesterase (ChE) activities of arthropod vectors of disease-causing agents exhibited various Michaelis-Menten KM values that were each lower than the KM value of bovine serum AChE. A lower KM value is indicative of higher affinity for substrate and is consistent with a hypothesized role in localized depletion of host tissue acetylcholine potentially modulating host immune responses at the arthropod bite site that may favor ectoparasite blood-feeding and alter host defensive responses against pathogen transmission and establishment.

Genotyping mutations in BmAChE3: A survey of organophosphate-resistant and -susceptible strains of Rhipicephalus (Boophilus) microplus.

Mutations I48L, I54V, R86Q, V137I, I492M, and T548A were identified previously in BmAChE3, a gene encoding acetylcholinesterase, from the organophosphate (OP) acaricide-resistant San Rommn strain of Rhipicephalus (Boophilus) microplus. Recombinant BmAChE3 acetylcholinesterase containing the R86Q mutation was shown to exhibit nearly 20-fold reduction in the rate of phosphorylation by paraoxon relative to the wild-type sequence. In addition, the R86Q mutation was present in resistant laboratory strains at elevated frequency compared with OP-susceptible strains but was insufficient to alone generate the OP-resistant phenotype (J. Med. Entomol. 44: 1013-1018). Here, we developed assays to genotype the remaining five mutations and evaluated frequency of all six BmAChE3 mutations in individual R. microplus ticks from laboratory and Mexican field-collected strains. We found a substantial number of individuals in known OP-susceptible strains that seemed to be homozygous for each of the mutations surveyed, the exception being I48L, which was infrequent in all strains, leading us to conclude that none of the mutations alone were responsible for generation of phenotypic resistance to OP acaricide.

Acetylcholinesterase mutation in diazinon-resistant Haematobia irritans (L.) (Diptera: Muscidae).

Acetylcholinesterase (AChE) cDNA from individual field-collected diazinon-resistant horn flies was amplified by RT-PCR. Sequencing of the amplification products revealed that 8/12 of the diazinon-resistant horn flies contained a point mutation previously associated with resistance to organophosphates in house flies and Drosophila, strongly suggesting that this cDNA encodes the AChE that is the target site for organophosphate (OP) pesticide. The point mutation (G262A) resulted in a shift from glycine to alanine in the mature HiAChE amino acid sequence at position 262. Allele-specific PCR and RLFP assays were developed to diagnose the presence or absence of the G262A mutation in individual flies. Use of the allele-specific assays each demonstrated the presence of the G262A mutation in 10 of 12 individual field-collected flies, demonstrating higher sensitivity than direct sequencing of RT-PCR amplification products. The G262A mutation was found in additional fly populations previously characterized as OP-resistant, further supporting that this AChE is the target site for OP pesticide. The allele-specific assay is a useful tool for quantitative assay of the resistance allele in horn fly populations.

TICK VECTOR AND DISEASE PATHOGEN SURVEILLANCE OF NILGAI ANTELOPE ( BOSELAPHUS TRAGOCAMELUS) IN SOUTHEASTERN TEXAS, USA.

Nilgai ( Boselaphus tragocamelus) are nonnative bovines that were originally introduced as game animals to one large, south Texas, US ranch but that are now present throughout southeastern Texas from Baffin Bay to Harlingen and in northern Mexico at least as far west as Durango. Between October 2014 and January 2017, nilgai ( n=517) were examined for the presence of tick ectoparasites, with particular interest in the cattle fever tick, Rhipicephalus ( Boophilus) microplus. These animals were either hunter killed or they were culled as part of federal cooperative harvesting from Cameron and Willacy counties in southeastern Texas. The proportion of fever tick-infested animals differed in a N-to-S pattern, and this was at least partly attributed to differences in habitat. The southern area is a lowland floodplain predominated by halophytes, whereas the northerly area is upland thorn scrub, the latter of which provides a vegetative canopy that is more conducive to tick survival and persistence. A subset of nilgai, all from the Texas-Mexico border area, were screened for livestock pathogens using molecular and serological assays. All nilgai were seronegative for Babesia ( Theileria) equi and Babesia cabalii. Although 11 animals were seropositive for Anaplasma marginale by competitive enzyme-linked immunosorbent assay (cELISA), these were interpreted with caution because of the lack of concordance between cELISA and molecular detection assays. All animals were PCR negative for presence of Babesia spp. DNA, and a single nilgai was seropositive for Babesia bovis and Babesia bigemina by complement fixation. It remains unknown whether cattle Babesia spp. can establish an infection in nilgai.

Fungi Isolated From House Flies (Diptera: Muscidae) on Penned Cattle in South Texas.

Musca domestica L. were collected from cattle diagnosed with bovine ringworm to evaluate the potential of the house fly to disseminate Trichophyton verrucosum E. Bodin, a fungal dermatophyte that is the causative agent for ringworm in cattle. Fungal isolates were cultured from 45 individual flies on supplemented Sabouraud dextrose agar, and isolates were identified using morphological and microscopic approaches. Each isolate was identified further by PCR amplification of the ribosomal DNA locus with fungal-specific primers and subsequent amplicon sequencing. Trichophyton verrucosum was not identified using these approaches. However, 35 different fungal species representing 17 genera were cultured from collected flies, including several species that are allergenic and pathogenic to humans and animals. Several species within the fungal orders Hypocreales, Microascales, Onygenales, Saccharomycetales, Xylaniales, and Agaricales were observed for the first time on house flies. The most frequent fungus recovered was Cladosporium cladosporoides Fresen, which is known to be a ubiquitous, airborne allergen to humans.

A Virulent Babesia bovis Strain Failed to Infect White-Tailed Deer (Odocoileus virginianus).

Wildlife are an important component in the vector-host-pathogen triangle of livestock diseases, as they maintain biological vectors that transmit pathogens and can serve as reservoirs for such infectious pathogens. Babesia bovis is a tick-borne pathogen, vectored by cattle fever ticks, Rhipicephalus spp., that can cause up to 90% mortality in naive adult cattle. While cattle are the primary host for cattle fever ticks, wild and exotic ungulates, including white-tailed deer (WTD), are known to be viable alternative hosts. The presence of cattle fever tick populations resistant to acaricides raises concerns regarding the possibility of these alternative hosts introducing tick-borne babesial parasites into areas free of infection. Understanding the B. bovis reservoir competence of these alternative hosts is critical to mitigating the risk of introduction. In this study, we tested the hypothesis that WTD are susceptible to infection with a B. bovis strain lethal to cattle. Two groups of deer were inoculated intravenously with either B. bovis blood stabilate or a larval extract supernatant containing sporozoites from infected R. microplus larvae. The collective data demonstrated that WTD are neither a transient host nor reservoir of B. bovis. This conclusion is supported by the failure of B. bovis to establish an infection in deer regardless of inoculum. Although specific antibody was detected for a short period in the WTD, the PCR results were consistently negative at multiple time points throughout the experiment and blood from WTD that had been exposed to parasite, transferred into naïve recipient susceptible calves, failed to establish infection. In contrast, naïve steers inoculated intravenously with either B. bovis blood stabilate or the larval extract supernatant containing sporozoites rapidly succumbed to disease. These findings provide evidence that WTD are not an epidemiological component in the maintenance of B. bovis infectivity to livestock.