scholarly journals Collecting Deer Keds (Diptera: Hippoboscidae: Lipoptena Nitzsch, 1818 and Neolipoptena Bequaert, 1942) and Ticks (Acari: Ixodidae) From Hunter-Harvested Deer and Other Cervids

2020 ◽  
Vol 20 (6) ◽  
Author(s):  
Karen C Poh ◽  
Michael Skvarla ◽  
Jesse R Evans ◽  
Erika T Machtinger
Keyword(s):  
Ixodes Scapularis ◽  
Blood Feeding ◽  
Host Choice ◽  
Sampling Scheme ◽  
Standardized Protocol ◽  
Anterior Dorsal ◽  
Collection Process ◽  
Geographical Regions ◽  
Materials Used ◽  
Anterior Posterior

Abstract Deer keds (Diptera: Hippoboscidae: Lipoptena Nitzsch, 1818 and Neolipoptena Bequaert, 1942) are blood-feeding ectoparasites that primarily attack cervids and occasionally bite humans, while ticks may be found on cervids, but are more generalized in host choice. Recent detection of pathogens such as Anaplasma and Borrelia in deer keds and historical infections of tick-borne diseases provides reason to investigate these ectoparasites as vectors. However, previous methods employed to sample deer keds and ticks vary, making it difficult to standardize and compare ectoparasite burdens on cervids. Therefore, we propose a standardized protocol to collect deer keds and ticks from hunter-harvested deer, which combines previous methods of sampling, including timing of collections, dividing sections of the deer, and materials used in the collection process. We tested a three-section and a five-section sampling scheme in 2018 and 2019, respectively, and found that dividing the deer body into five sections provided more specificity in identifying where deer keds and ticks may be found on deer. Data from 2018 suggested that deer keds and ticks were found on all three sections (head, anterior, posterior), while data from 2019 suggested that more Ixodes scapularis were found on the head and deer keds were found on all body sections (head, dorsal anterior, dorsal posterior, ventral anterior, and ventral posterior). The protocol provides an efficient way to sample deer for deer keds and ticks and allows researchers to compare ectoparasite burdens across geographical regions. Furthermore, this protocol can be used to collect other ectoparasites from deer or other cervids.

scholarly journals Genetic Analysis of Powassan virus Genotypes Circulating in North America [University of Findlay]

2019 ◽  
Author(s):  
Kristen Huseman ◽  
Abby L Levitt
Keyword(s):  
North America ◽  
Host Species ◽  
Evolutionary Dynamics ◽  
Genetic Recombination ◽  
Ixodes Scapularis ◽  
Geographic Location ◽  
Recombination Analysis ◽  
Phenotypic Characters ◽  
Geographical Regions ◽  
Powassan Virus

Powassan virus (POWV), a tick-borne flavivirus, is the only member of the tick-borne encephalitis serogroup found in North America. Two distinct lineages, prototype lineage (POWV, lineage I) and deer tick virus (DVT, lineage II) are maintained in enzootic transmission cycles by different tick species based on different geographical regions. In North America, Ixodes scapularis acts as the primary vector throughout the Northeast, and Ixodes cookei throughout the Midwest and much of Canada. Importantly, the incidence of human disease due to POWV has increased by 671% over the last 18 years; with DVT perhapes being the most worrisome genotype due to the wide spread range and prevalence of the vector species. The aim of this study is to assess the evolutionary dynamics of the POWV/DTV complex using the most current full-genome, envelope, and 3’UTR sequences available. Bayesian phylogenetic inferences support the two distinct, monophyletic lineages corresponding to POWV and DTV. Additional analysis were performed in order to quantify the degree to which viral phenotypic characters (such as geographic location, and host species utilization) are correlated with shared ancestry within the two different genotypes. The results of this phylogeny-trait correlation analysis suggest significant clustering of viral sequence by sample location. Such in situ evolution is compatible with the relatively limited distances traveled by most ticks and their prefered host species. Lastly, genetic recombination analysis shows no evidence of between genotype recombination, however further analysis needs to be conducted using within genotype recombination analysis as recombination events may be restricted to only closely genetically related viruses.

scholarly journals Ixodes scapularis Tick Saliva Proteins Sequentially Secreted Every 24 h during Blood Feeding

2016 ◽  
Vol 10 (1) ◽  
pp. e0004323 ◽  
Author(s):  
Tae Kwon Kim ◽  
Lucas Tirloni ◽  
Antônio F. M. Pinto ◽  
James Moresco ◽  
John R. Yates ◽  
...  
Keyword(s):  
Ixodes Scapularis ◽  
Blood Feeding ◽  
Tick Saliva

scholarly journals Wide Dispersal and Possible Multiple Origins of Low-Copy-Number Plasmids in Rickettsia Species Associated with Blood-Feeding Arthropods

2010 ◽  
Vol 76 (6) ◽  
pp. 1718-1731 ◽  
Author(s):  
Gerald D. Baldridge ◽  
Nicole Y. Burkhardt ◽  
Marcelo B. Labruna ◽  
Richard C. Pacheco ◽  
Christopher D. Paddock ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Ixodes Scapularis ◽  
Blood Feeding ◽  
Adaptive Traits ◽  
Phylogenetic Groups ◽  
Plasmid Copy ◽  
Rickettsia Species ◽  
Real Time Quantitative Pcr ◽  
Rickettsia Amblyommii ◽  
Obligate Intracellular Bacteria

ABSTRACT Plasmids are mobile genetic elements of bacteria that can impart important adaptive traits, such as increased virulence or antibiotic resistance. We report the existence of plasmids in Rickettsia (Rickettsiales; Rickettsiaceae) species, including Rickettsia akari, “Candidatus Rickettsia amblyommii,” R. bellii, R. rhipicephali, and REIS, the rickettsial endosymbiont of Ixodes scapularis. All of the rickettsiae were isolated from humans or North and South American ticks. R. parkeri isolates from both continents did not possess plasmids. We have now demonstrated plasmids in nearly all Rickettsia species that we have surveyed from three continents, which represent three of the four major proposed phylogenetic groups associated with blood-feeding arthropods. Gel-based evidence consistent with the existence of multiple plasmids in some species was confirmed by cloning plasmids with very different sequences from each of two “Ca. Rickettsia amblyommii” isolates. Phylogenetic analysis of rickettsial ParA plasmid partitioning proteins indicated multiple parA gene origins and plasmid incompatibility groups, consistent with possible multiple plasmid origins. Phylogenetic analysis of potentially host-adaptive rickettsial small heat shock proteins showed that hsp2 genes were plasmid specific and that hsp1 genes, found only on plasmids of “Ca. Rickettsia amblyommii,” R. felis, R. monacensis, and R. peacockii, were probably acquired independently of the hsp2 genes. Plasmid copy numbers in seven Rickettsia species ranged from 2.4 to 9.2 per chromosomal equivalent, as determined by real-time quantitative PCR. Plasmids may be of significance in rickettsial evolution and epidemiology by conferring genetic plasticity and host-adaptive traits via horizontal gene transfer that counteracts the reductive genome evolution typical of obligate intracellular bacteria.

scholarly journals Blood Digestion by Trypsin-Like Serine Proteases in the Replete Lyme Disease Vector Tick, Ixodes scapularis

Insects ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 201
Author(s):  
Jeremiah Reyes ◽  
Cuauhtemoc Ayala-Chavez ◽  
Arvind Sharma ◽  
Michael Pham ◽  
Andrew B. Nuss ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Blood Meal ◽  
Serine Proteases ◽  
Ixodes Scapularis ◽  
Life Stage ◽  
Blood Feeding ◽  
Eastern United States ◽  
Blood Digestion ◽  
Hemoglobin Degradation

Ixodes scapularis is the major vector of Lyme disease in the Eastern United States. Each active life stage (larva, nymph, and adult) takes a blood meal either for developing and molting to the next stage (larvae and nymphs) or for oviposition (adult females). This protein-rich blood meal is the only food taken by Ixodes ticks and therefore efficient blood digestion is critical for survival. Studies in partially engorged ticks have shown that the initial stages of digestion are carried out by cathepsin proteases within acidic digestive cells. In this study, we investigated the potential role of serine proteases in blood digestion in replete ticks. RNA interference was used for functional analysis and a trypsin-benzoyl-D, L-arginine 4-nitoanilide assay was used to measure active trypsin levels. Hemoglobinolytic activity was determined in vitro, with or without a serine protease inhibitor. Our data suggest that trypsin levels increase significantly after repletion. Knockdown of serine proteases negatively impacted blood feeding, survival, fecundity, levels of active trypsin in the midgut, and resulted in lower hemoglobin degradation. Incubation of midgut extract with a trypsin inhibitor resulted in 65% lower hemoglobin degradation. We provide evidence of the serine proteases as digestive enzymes in fully engorged, replete females. Understanding the digestive profile of trypsin during blood meal digestion in I. scapularis improves our understanding of the basic biology of ticks and may lead to new methods for tick control.

scholarly journals Assessing the blood-host plasticity and dispersal rate of the malaria vector Anopheles coluzzii

2018 ◽  
Author(s):  
James Orsborne ◽  
Luis Furuya-Kanamori ◽  
Claire L. Jeffries ◽  
Mojca Kristan ◽  
Abdul Rahim Mohammed ◽  
...  
Keyword(s):  
Blood Meal ◽  
Disease Transmission ◽  
Blood Feeding ◽  
Host Choice ◽  
Anopheles Coluzzii ◽  
Dispersal Rate ◽  
Blood Digestion ◽  
Novel Method ◽  
Blood Meal Digestion ◽  
Blood Meals

AbstractDifficulties with observing the dispersal of insect vectors in the field have hampered understanding of several aspects of their behaviour linked to disease transmission. Here, a novel method based on detection of blood-meal sources is introduced to inform two critical and understudied mosquito behaviours: plasticity in the malaria vector’s blood-host choice and vector dispersal. Strategically located collections of Anopheles coluzzii from a malaria-endemic village of southern Ghana showed statistically significant variation in host species composition of mosquito blood-meals. Trialling a new sampling approach gave the first estimates for the remarkably local spatial scale across which host choice is plastic. Using quantitative PCR, the blood-meal digestion was then quantified for field-caught mosquitoes and calibrated according to timed blood digestion in colony mosquitoes. We demonstrate how this new ‘molecular Sella score’ approach can be used to estimate the dispersal rate of blood-feeding vectors caught in the field.

scholarly journals The Phylogenetic Analysis of Cimex hemipterus (Hemiptera: Cimicidae) Isolated from Different Regions of Iran Using Cytochrome Oxidase Subunit I Gene

2020 ◽  
Author(s):  
Awat Samiei ◽  
Mousa Tavassoli ◽  
Karim Mardani
Keyword(s):  
Dna Sequencing ◽  
Cytochrome Oxidase ◽  
Blood Feeding ◽  
Coi Gene ◽  
Sequencing Analysis ◽  
Nucleotide Polymorphisms ◽  
Bed Bugs ◽  
Geographical Regions ◽  
Dna Sequencing Analysis ◽  
I Gene

Background: Bedbugs are blood feeding ectoparasites of humans and several domesticated animals. There are scar­city of information about the bed bugs population throughout Iran and only very limited and local studies are availa­ble. The aim of this study is to assess the phylogenetic relationships and nucleotide diversity using partial sequences of cytochrome oxidase I gene (COI) among the populations of tropical bed bugs inhabiting Iran. Methods: The bedbugs were collected from cities located in different geographical regions of Iran. After DNA ex­traction PCR was performed for COI gene using specific primers. Then DNA sequencing was performed on PCR products for the all 15 examined samples. Results: DNA sequencing analysis showed that the all C. hemipterus samples were similar, despite the minor nu­cleotide variations (within the range of 576 to 697bp) on average between 5 and 10 Single nucleotide polymorphisms (SNPs). Subsequently, the results were compared with the database in gene bank which revealed close similarity and sequence homology with other C. hemipterus from other parts of the world. Conclusion: In conclusion, this study has demonstrated the ability of the COI gene to differentiate between the C. hemipterus populations from a few different locations in Iran. The current research is the first report of phylogenetic and genetic species diversity analysis conducted on C. hemipterus in Iran. These results provided basic information for further studies of molecular epidemiology, public health and pest control operators in Iran.

The anterior extent of dorsal development of the Xenopus embryonic axis depends on the quantity of organizer in the late blastula

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 363-372 ◽  
Author(s):  
R.M. Stewart ◽  
J.C. Gerhart
Keyword(s):  
Xenopus Laevis ◽  
Cell Population ◽  
Marginal Zone ◽  
Organizer Region ◽  
Blastula Stage ◽  
Dorsal Midline ◽  
Anterior Dorsal ◽  
Lateral Width ◽  
Anterior Extent ◽  
Anterior Posterior

In amphibian gastrulae, the cell population of the organizer region of the marginal zone (MZ) establishes morphogenesis and patterning within itself and within surrounding regions of the MZ, presumptive neurectoderm, and archenteron roof. We have tested the effects on pattern of reducing the amount of organizer region by recombining halves of Xenopus laevis late blastulae cut at different angles from the bilateral plane. When regions within 30 degrees of the dorsal midline are excluded from recombinants, ventralized embryos develop lacking the entire anterior-posterior sequence of dorsal structures, suggesting that the organizer is only 60 degrees wide (centered on the dorsal midline) at the late blastula stage. As more and more dorsal MZ (organizer) is included in the recombinant, progressively more anterior dorsal structures are formed. In all cases, when any dorsal structures are missing they are deleted serially from the anterior end. Thus, we suggest that the amount (lateral width) of the organizer in the MZ determines the anterior extent of dorsal development.

scholarly journals Blood Digestion by Trypsin-Like Serine Protease in the Replete Lyme Disease Vector Tick, Ixodes scapularis

2020 ◽  
Author(s):  
Jeremiah Reyes ◽  
Cuauhtemoc Ayala-Chavez ◽  
Michael Pham ◽  
Arvind Sharma ◽  
Andrew Nuss ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Serine Protease ◽  
Blood Meal ◽  
Serine Proteases ◽  
Ixodes Scapularis ◽  
Life Stage ◽  
Blood Feeding ◽  
Blood Digestion ◽  
Hemoglobin Degradation

Ixodes scapularis is the major vector of Lyme disease in the eastern United States. Each active life stage (larva, nymph, and adult) takes a blood meal either for developing and molting to the next stage (larvae and nymphs) or for oviposition (adult females). This protein-rich blood meal is the only food taken by Ixodes ticks and therefore blood digestion is very important for tick survival. Most studies on blood digestion in ticks have shown that the initial stages of digestion are carried out by cathepsin proteases within acidic digestive cells. However, most of these studies have focused on partially engorged ticks. In other hematophagous arthropods, the serine proteases play an important role in blood protein degradation. In this study, we determined transcript expression of four I. scapularis serine proteases with previously characterized roles in blood digestion. RNA interference was used for functional analysis and a trypsin-benzoyl-D, L-arginine 4-nitoanilide assay was used to measure active trypsin levels. An in vitro hemoglobinolytic assay was performed with or without serine protease inhibitor. Our data suggest that trypsin levels increase significantly after blood feeding and peaked in larvae, nymphs, and adults at 3, 1, and 1 day post host detachment, respectively. The knockdown of three previously identified serine proteases by RNAi negatively impacted blood intake, survival, fecundity, levels of active trypsin in the gut and resulted in lower hemoglobin degradation in vitro. A trypsin inhibitor, PMSF, blocked the action of trypsin in the gut extract resulting in 65% lower hemoglobin degradation. We provide evidence of the serine proteases as digestive enzymes in fully engorged, replete females. Our data also demonstrated that in addition to blood digestion, these serine proteases might have a role in blood feeding success in I. scapularis.

scholarly journals Selective human cysteine protease inhibition mediates Ixodes scapularis blood feeding success

2008 ◽  
Vol 22 (S1) ◽  
Author(s):  
Michail Kotsyfakis ◽  
Shahid Karim ◽  
Jennifer Anderson ◽  
John Andersen ◽  
Jesus Valenzuela ◽  
...  
Keyword(s):  
Cysteine Protease ◽  
Ixodes Scapularis ◽  
Blood Feeding ◽  
Protease Inhibition ◽  
Feeding Success

scholarly journals Blood Digestion by Trypsin-Like Serine Protease in the Lyme Disease Vector Tick, Ixodes Scapularis

2019 ◽  
Author(s):  
Jeremiah Reyes ◽  
Cuauhtemoc Ayala-Chavez ◽  
Andrew Nuss ◽  
Monika Gulia-Nuss
Keyword(s):  
Lyme Disease ◽  
Serine Protease ◽  
Blood Meal ◽  
Ixodes Scapularis ◽  
Life Stage ◽  
Blood Feeding ◽  
Eastern United States ◽  
Blood Digestion ◽  
Early Protein ◽  
Gut Ph

Ixodes scapularis is the major vector of Lyme disease in the eastern United States. This species undergoes a life cycle consisting of eggs and three active stages: larva, nymph, and adult. Each active life stage takes a blood meal either for developing and molting to the next stage (larvae and nymphs) or for oviposition (adult females). This protein rich blood meal is the only food taken by Ixodes ticks. Most studies on blood digestion in ticks have shown that the initial stages of blood digestion are carried out by cathepsin proteases within endosomes of acidic digestive cells. However, in other hematophagous arthropods, the serine protease trypsin plays an important role in early protein degradation. In this study, we determined transcript expression of I. scapularis cathepsins and serine proteases, some with previously characterized roles in blood digestion. Gut pH was also determined and a trypsin-benzoyl-D, L-arginine 4-nitoanilide assay was used to measure active trypsin levels during blood digestion. Our data suggest that trypsin levels increase significantly after blood feeding and peaked in larvae, nymphs, and adults at 3, 1, and 1 days post host detachment, respectively. In addition, alkaline gut pH (8.0) conditions after I. scapularis blood feeding were similar to those required for trypsin activity in other arthropods suggesting these enzymes have an important and previously overlooked role in I. scapularis blood digestion.

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