Resistance of Nguni, Bonsmara and Hereford cattle to ticks in a Bushveld region of South Africa

Counts of engorged female ticks on naturally infested cattle over a 2 year period, showed that indigenous Nguni cattle harboured significantly fewer Amblyomma hebraeum, Boophilus decoloratus and Hyalomma spp. during periods of peak abundance than either Bonsmara or Hereford cattle. Fewer abscesses, associated with tick bite, were also present in the Nguni cattle. Individual tick resistance indices, determined after artificial tick infestation in the field, could not be correlated with hair length, skin thickness or conglutinin titres. The consistently large percentage of Nguni cattle showing high tick resistance according to index determinations, indicates a superior level of natural immunity in this breed. The relative incidence of individuals in high, medium and low resistance classes reflected an increase in resistance with exposure to ticks and the potential for the selection for tick resistance within all 3 breeds.The articles have been scanned in colour with a HP Scanjet 5590; 600dpi. Adobe Acrobat XI Pro was used to OCR the text and also for the merging and conversion to the final presentation PDF-format.lmchunu2014mn201

The host preferences of Nuttalliella namaqua (Ixodoidea : Nuttalliellidae) : a generalist approach to surviving multiple host-switches

Nuttalliella namaqua has been described as a “living fossil” and the closest extant species to the ancestral tick lineage. It was previously proposed that the Nuttalliella lineage parasitized reptile-like mammals in the Permian and had to switch hosts several times due to mass or host lineage extinctions. Extant hosts include girdled lizards and murid rodents. The current study extends knowledge on the extant host range of N. namaqua using gut meal analysis of field collected specimens. Nymphs and females can parasitize a variety of reptiles that includes skinks, geckos and girdled lizards. Blood-meal from a hyrax was also detected in a specimen suggesting that N. namaqua could parasitize a broader range of mammals than the previously suggested murid rodents. Rather than being host specific, N. namaqua is proposed to be a generalist and the ability to switch and parasitize multiple hosts allowed it to survive multiple mass and host lineage extinctions.The Joy Liebenberg Trust (21/19/JT02) allocated to BM, an incentive funding for rated researchers grant from the National Research Foundation of South Africa (NRF-Mans 76499) and a South African National Research Foundation grant allocated to AL (NRF-Spain).http://link.springer.com/journal/10493hb2014ab201

Next-generation sequencing as means to retrieve tick systematic markers, with the focus on Nuttalliella namaqua (Ixodoidea : Nuttalliellidae)

Nuclear ribosomal RNA (18S and 28S rRNA) and mitochondrial genomes are commonly used in tick systematics. The ability to retrieve these markers using next-generation sequencing was investigated using the tick Nuttalliella namaqua. Issues related to nuclear markers may be resolved using this approach, notably, the monotypic status of N. namaqua and its basal relationship to other tick families. Four different Illumina datasets (∼55 million, 100 bp reads each) were generated from a single tick specimen and assembled to give 350k-390k contigs. A genome size of ∼1 Gbp was estimated with low levels of repetitive elements. Contigs (>1000 bp, >50-fold coverage) present in most assemblies (n=69), included host-derived 18S and 28S rRNA, tick and host-derived transposable elements, full-length tick 18S and 28 rRNA, the mitochondrial genome in single contig assemblies and the histone cassette. Coverage for the nuclear rRNA genes was above 1000-fold confirming previous sequencing errors in the 18S rRNA gene, thereby maintaining the monotypic status of this tick. Nuclear markers for the soft tick Argas africolumbae were also retrieved from next-generation data. Phylogenetic analysis of a concatenated 18S-28S rRNA dataset supported the grouping of N. namaqua at the base of the tick tree and the two main tick families in separate clades. This study confirmed the monotypic status of N. namaqua and its basal relationship to other tick families. Next-generation sequencing of genomic material to retrieve high quality nuclear and mitochondrial systematic markers for ticks is viable and may resolve issues around conventional sequencing errors when comparing closely related tick species.The Joy Liebenberg Trust (21/19/JT02) allocated to BM, incentive funding for rated researchers grant from the National Research Foundation of South Africa (NRF-Mans 76499), and the ARC Tick Vaccine project (30/01/V010).http://www.elsevier.com/locate/ttbdishb2017Veterinary Tropical Disease

An update of the tsetse fly (Diptera: Glossinidae) distribution and African animal trypanosomosis prevalence in north-eastern KwaZulu-Natal, South Africa

An unpredicted outbreak of African animal trypanosomosis or nagana in 1990 in north-eastern KwaZulu-Natal necessitated an emergency control programme, utilising the extensive cattledipping system in the area, as well as a reassessment of the tsetse and trypanosomosis problem in the province. Since 1990, sporadic blood sampling of cattle at the dip tanks in the naganainfested areas were undertaken to identify trypanosome species involved and to determine the infection prevalence in cattle. The distribution and species composition of the tsetse populations in the area were also investigated. From November 2005 to November 2007 selected dip tanks were surveyed for trypanosome infection prevalence. During April 2005 to August 2009 the distribution and abundance of tsetse populations were assessed with odour-baited H traps. The tsetse and trypanosome distribution maps were updated and potential correlations between tsetse apparent densities (ADs) and the prevalence of trypanosomosis were assessed. Glossina brevipalpis Newstead and Glossina austeni Newstead were recorded in locations where they have not previously been collected. No significant correlation between tsetse relative abundance and nagana prevalence was found, which indicated complex interactions between tsetse fly presence and disease prevalence. This was epitomised by data that indicated that despite large differences in the ADs of G. austeni and G. brevipalpis, trypanosome infection prevalence was similar in all three districts in the area. This study clearly indicated that both tsetse species play significant roles in trypanosome transmission and that it will be essential that any control strategy, which aims at sustainable management of the disease, should target both species

The Mitochondrial genomes of Nuttalliella namaqua (Ixodoidea: Nuttalliellidae) and Argas africolumbae (Ixodoidae: Argasidae) : estimation of divergence dates for the major tick lineages and reconstruction of ancestral blood-feeding characters

Ixodida are composed of hard (Ixodidae), soft (Argasidae) and the monotypic Nuttalliellidae (Nuttalliella namaqua) tick families. Nuclear 18S rRNA analysis suggested that N. namaqua was the closest extant relative to the last common ancestral tick lineage. The mitochondrial genomes of N. namaqua and Argas africolumbae were determined using next generation sequencing and de novo assembly to investigate this further. The latter was included since previous estimates on the divergence times of argasids lacked data for this major genus. Mitochondrial gene order for both was identical to that of the Argasidae and Prostriata. Bayesian analysis of the COI, Cytb, ND1, ND2 and ND4 genes confirmed the monophyly of ticks, the basal position of N. namaqua to the other tick families and the accepted systematic relationships of the other tick genera. Molecular clock estimates were derived for the divergence of the major tick lineages and supported previous estimates on the origins of ticks in the Carboniferous. N. namaqua larvae fed successfully on lizards and mice in a prolonged manner similar to many argasids and all ixodids. Excess blood meal-derived water was secreted via the salivary glands, similar to ixodids. We propose that this prolonged larval feeding style eventually gave rise to the long feeding periods that typify the single larval, nymphal and adult stages of ixodid ticks and the associated secretion of water via the salivary glands. Ancestral reconstruction of characters involved in blood-feeding indicates that most of the characteristics unique to either hard or soft tick families were present in the ancestral tick lineage.The Joy Liebenberg Trust (21/19/JT02) allocated to BM and a South African National Research Foundation grant allocated to AL (NRF-Spain).http://www.plosone.orgam2013ab201

Nuttalliella namaqua (Ixodoidea: Nuttalliellidae) : first description of the male, immature stages and re-description of the female

Nuttalliella namaqua is the only species of the enigmatic third tick family. Females possess features of hard and soft ticks and have been designated as the ‘‘missing link’’ between the main tick families. Its position at the base of the tick tree suggests that some of the features unique to hard and soft ticks were present in the ancestral tick lineage. Larvae, nymphae and males have not been described to date and questions regarding their biological affinities to the main tick families remain unclear. The current study addressed these questions via the description of larvae, nymphae and males and resolved issues pertaining to female morphology. Field collected as well as laboratory-engorged females laid eggs and viable larvae subsequently hatched. The larvae possess morphological structures not present in subsequent stages: namely, a sclerotized scutum, pores on the dorsal surface of legs and a dentate anal plate. The last two characters are not present in ixodids and argasids. N. namaqua larvae and nymphae show a similar morphology to females: a unique hypostomal structure i.e., bluntly rounded apically in nymphae and females and ball-like in the larvae. A re-description of some structures in female N. namaqua has resolved differences in the original descriptions, namely that N. namaqua have 4 palpal segments as found in ixodids and argasids and posthypostomal setae. The male was discovered for the first time and described. Characteristic male features include: a pseudoscutum over most of the dorsum, an outgrowth on the chelicerae forming a unique rod-like structure similar to a spematodactyl in mites and medial extension of palpal segment 2 forming a large ventral crib for segment 4. All life stages possess some features found in hard and soft ticks and its status as the ‘‘missing link’’ between the tick families remains.The Joy Liebenberg Trust (21/19/JT02) allocated to BM, a South African National Research Foundation grant allocated to AL (NRF-Spain).http://www.plosone.orgab201

De novo assembly and annotation of the salivary gland transcriptome of Rhipicephalus appendiculatus male and female ticks during blood feeding

Tick secretory proteins modulate haemostasis, inflammation and immune responses of the host and are attractive recombinant anti-tick vaccine candidates. Yet, many of the proteins have not been characterised due to the limited sequence availability for ticks and other arthropods for homology-based annotation. To address this limitation, we sequenced the salivary glands of the economically important adult male and female Rhipicephalus appendiculatus ticks during feeding. The quality-filtered Illumina sequencing reads were de novo assembled to generate a R. appendiculatus sialotranscriptome of 21,410 transcripts. A non-redundant set of 12,761 R. appendiculatus proteins was predicted from the transcripts, including 2134 putative secretory and 8237 putative housekeeping proteins. Secretory proteins accounted for most of the expression in the salivary gland transcriptome (63%). Of the secretory protein class, the Glycine-rich superfamily contributed 66% and the Lipocalin family 12% of the transcriptome expression. Differential expression analysis identified 1758 female and 2346 male up-regulated transcripts, suggesting varying blood-feeding mechanisms employed between female and male ticks. The sialotranscriptome assembled in this work, greatly improves on the sequence information available for R. appendiculatus and is a valuable resource for potential future vaccine candidate selection.The Economic Competitive Support Programme (30/01/V010) and Incentive Funding for Rated Researchers (NRF-Mans).http://www.elsevier.com/locate/ttbdis2017-06-30hb2016Veterinary Tropical Disease

Metallated phthalocyanines and their hydrophilic derivatives for multi-targeted oncological photodynamic therapy

Background and aim: A photosensitizer (PS) delivery and comprehensive tumor targeting platform was developed that is centered on the photosensitization of key pharmacological targets in solid tumors (cancer cells, tumor vascular endothelium, and cellular and non-cellular components of the tumor microenvironment) before photodynamic therapy (PDT). Interstitially targeted liposomes (ITLs) encapsulating zinc phthalocyanine (ZnPC) and aluminum phthalocyanine (AlPC) were formulated for passive targeting of the tumor microenvironment. In previous work it was established that the PEGylated ITLs were taken up by cultured cholangiocarcinoma cells. The aim of this study was to verify previous results in cancer cells and to determine whether the ITLs can also be used to photosensitize cells in the tumor microenvironment and vasculature. Following positive results, rudimentary in vitro and in vivo experiments were performed with ZnPC-ITLs and AlPC-ITLs as well as their water-soluble tetrasulfonated derivatives (ZnPCS4 and AlPCS4) to assemble a research dossier and bring this platform closer to clinical transition. Methods: Flow cytometry and confocal microscopy were employed to determine ITL uptake and PS distribution in cholangiocarcinoma (SK-ChA-1) cells, endothelial cells (HUVECs), fibroblasts (NIH-3T3), and macrophages (RAW 264.7). Uptake of ITLs by endothelial cells was verified under flow conditions in a flow chamber. Dark toxicity and PDT efficacy were determined by cell viability assays, while the mode of cell death and cell cycle arrest were assayed by flow cytometry. In vivo systemic toxicity was assessed in zebrafish and chicken embryos, whereas skin phototoxicity was determined in BALB/c nude mice. A PDT efficacy pilot was conducted in BALB/c nude mice bearing human triple-negative breast cancer (MDA-MB-231) xenografts. Results: The key findings were that (1) photodynamically active PSs (i.e., all except ZnPCS4) were able to effectively photosensitize cancer cells and non-cancerous cells; (2) following PDT, photodynamically active PSs were highly toxic-to-potent as per anti-cancer compound classification; (3) the photodynamically active PSs did not elicit notable systemic toxicity in zebrafish and chicken embryos; (4) ITL-delivered ZnPC and ZnPCS4 were associated with skin phototoxicity, while the aluminum-containing PSs did not exert detectable skin phototoxicity; and (5) ITL-delivered ZnPC and AlPC were equally effective in their tumor-killing capacity in human tumor breast cancer xenografts and superior to other non-phthalocyanine PSs when appraised on a per mole administered dose basis. Conclusions: AlPC(S4) are the safest and most effective PSs to integrate into the comprehensive tumor targeting and PS delivery platform. Pending further in vivo validation, these third-generation PSs may be used for multi-compartmental tumor photosensitization

Amplification and sequencing of entire tick mitochondrial genomes for a phylogenomic analysis

The mitochondrial genome (mitogenome) has proven to be important for the taxonomy, systematics, and population genetics of ticks. However, current methods to generate mitogenomes can be costprohibitive at scale. To address this issue, we developed a cost-effective approach to amplify and sequence the whole mitogenome of individual tick specimens. Using two different primer sites, this approach generated two full-length mitogenome amplicons that were sequenced using the Oxford Nanopore Technologies’ Mk1B sequencer. We used this approach to generate 85 individual tick mitogenomes from samples comprised of the three tick families, 11 genera, and 57 species. Twentysix of these species did not have a complete mitogenome available on GenBank prior to this work. We benchmarked the accuracy of this approach using a subset of samples that had been previously sequenced by low-coverage Illumina genome skimming. We found our assemblies were comparable or exceeded the Illumina method, achieving a median sequence concordance of 99.98%. We further analyzed our mitogenome dataset in a mitophylogenomic analysis in the context of all three tick families. We were able to sequence 72 samples in one run and achieved a cost/sample of ~ $10 USD. This cost-effective strategy is applicable for sample identification, taxonomy, systematics, and population genetics for not only ticks but likely other metazoans; thus, making mitogenome sequencing equitable for the wider scientific community.NIH Grants and the Norman E. Borlaug International Agricultural Science and Technology Fellow.http://www.nature.com/scientificreportsam2023Veterinary Tropical Disease

The TT genotype of methylenetetrahydrofolate reductase 677C>T polymorphism increases the susceptibility to pediatric ischemic stroke: meta-analysis of the 822 cases and 1,552 controls

The 677C>T polymorphism within methylenetetrahydrofolate reductase (MTHFR) gene is related to an elevated level of homocysteine. Thus it may be considered as a genetic risk factor in ischemic stroke. Apparently studies of this type of polymorphism in childhood stroke have shown conflicting results. We performed meta-analysis of all the data that are available in relation with MTHFR polymorphism and the risk of ischemic stroke in children. We searched PubMed (last search dated December 2010) using “MTHFR polymorphism”, “ischemic stroke” “child”, “children”, “pediatric stroke” as keywords and reference lists of studies and reviews on the topic. Finally, 15 case–control studies corresponded to the inclusion criteria for meta-analysis. These studies involved the total number of 822 children and adolescents after ischemic stroke and 1,552 control subjects. Fixed or random effects models were used depending on the heterogeneity between the studies. The association between ischemic stroke and 677C>T polymorphism within MTHFR gene was observed in three of the studies. The pooled analysis showed that TT genotype of MTHFR gene is more common in stroke patients than in controls (p = 0.0402, odds ratio = 1.57, 95 % confidence interval 1.02–2.41). The Egger’s test did not reveal presence of a publication bias. The results based on a sizeable group of cases and controls have proved that the 677C>T polymorphism in MTHFR gene is associated with the development of ischemic stroke in children