40 research outputs found
Comparative proteomic analysis of Neisseria meningitidis wildtype and dprA null mutant strains links DNA processing to pilus biogenesis
No full textBackground
DNA processing chain A (DprA) is a DNA binding protein which is ubiquitous in bacteria, and is required for DNA transformation to various extents among bacterial species. However, the interaction of DprA with competence and recombination proteins is poorly understood. Therefore, the proteomes of whole Neisseria meningitidis (Nm) wildtype and dprA mutant cells were compared. Such a comparative proteomic analysis increases our understanding of the interactions of DprA with other Nm components and may elucidate its potential role beyond DNA processing in transformation.
Results
Using label-free quantitative proteomics, a total of 1057 unique Nm proteins were identified, out of which 100 were quantified as differentially abundant (P ≤ 0.05 and fold change ≥ |2|) in the dprA null mutant. Proteins involved in homologous recombination (RecA, UvrD and HolA), pilus biogenesis (PilG, PilT1, PilT2, PilM, PilO, PilQ, PilF and PilE), cell division, including core energy metabolism, and response to oxidative stress were downregulated in the Nm dprA null mutant. The mass spectrometry data are available via ProteomeXchange with identifier PXD006121. Immunoblotting and co-immunoprecipitation were employed to validate the association of DprA with PilG. The analysis revealed reduced amounts of PilG in the dprA null mutant and reduced amounts of DprA in the Nm pilG null mutant. Moreover, a number of pilus biogenesis proteins were shown to interact with DprA and /or PilG.
Conclusions
DprA interacts with proteins essential for Nm DNA recombination in transformation, pilus biogenesis, and other functions associated with the inner membrane. Inverse downregulation of Nm DprA and PilG expression in the corresponding mutants indicates a link between DNA processing and pilus biogenesis
Effect of Brucella Infection on Reproduction Conditions of Female Breeding Cattle and Its Public Health Significance in Western Tigray, Northern Ethiopia
No full textThe most common clinical manifestations of brucellosis in livestock are associated with reproduction. This paper reports the result of a cross-sectional study conducted between October, 2007 and April, 2008 in Western Tigray, North Ethiopia, with the objectives of assessing the effect of Brucella infection on reproduction conditions of female breeding bovine and to explore the presence of Brucella seroreactors in vulnerable humans. A total of 1,354 and 246 sera were collected from female cattle and humans, respectively. The sera were screened using Rose Bengal plate test (RBPT), and positive samples were confirmed by complement fixation test (CFT). Reproductive conditions for female cattle and risk to human brucellosis seropositivity were tested by using logistic regression analysis. The result indicated that the overall prevalence in female cattle was 6.1%. The study showed 1.2% prevalence among human risk groups, all of which were herdsmen. Logistic regression identified parity status, calving interval, abortion history, and abortion period were significantly associated with seropositivity. The association was not significant with reproductive status and parity number. Proper hygienic practices and team work between veterinary and health personnel should improve the efforts to combat disease transmission
Tsetse bloodmeal analyses incriminate the common warthog Phacochoerus africanus as an important cryptic host of animal trypanosomes in smallholder cattle farming communities in Shimba Hills, Kenya
No full textTable S1: Data on tsetse fly bloodmeal hosts and trypanosome
infections in the different study-blocks in Shimba Hills, Kenya.Data Availability Statement: The dataset used and/or analysed during the current study are available from the corresponding author FIE on reasonable request. DNA sequences of vertebrate
species generated during the current study are available in the GenBank under accession numbers:
MZ816958-MZ816971.Trypanosomes are endemic and retard cattle health in Shimba Hills, Kenya. Wildlife in the area act as reservoirs of the parasites. However, wild animal species that harbor and expose cattle to tsetse-borne trypanosomes are not well known in Shimba Hills. Using xeno-monitoring surveillance to investigate wild animal reservoirs and sources of trypanosomes in Shimba Hills, we screened 696 trypanosome-infected and uninfected tsetse flies for vertebrate DNA using multiplegene PCR-High Resolution Melting analysis and amplicon sequencing. Results revealed that tsetse flies fed on 13 mammalian species, preferentially Phacochoerus africanus (warthogs) (17.39%, 95% CI: 14.56–20.21) and Bos taurus (cattle) (11.35%, 95% CI: 8.99–13.71). Some tsetse flies showed positive cases of bloodmeals from multiple hosts (3.45%, 95% CI: 2.09–4.81), including warthog and cattle (0.57%, 95% CI: 0.01–1.14). Importantly, tsetse flies that took bloodmeals from warthog had significant risk of infections with Trypanosoma vivax (5.79%, 95% CI: 1.57–10.00), T. congolense (7.44%, 95% CI: 2.70–12.18), and T. brucei sl (2.48%, 95% CI: 0.33–5.29). These findings implicate warthogs as important reservoirs of tsetse-borne trypanosomes affecting cattle in Shimba Hills and provide valuable epidemiological insights to underpin the parasites targeted management in Nagana vector control programs in the area.A German Academic Exchange Service (DAAD) in-region postgraduate scholarship; the BioVision Foundation Switzerland; European Union’s Integrated Biological Control Applied Research Programme—tsetse repellent component; the German Ministry for Economic Cooperation and Development (BMZ) through the Deutsche Gesellschaft für Internationale Zusammenarbeit; UK’s Department for International Development (DFID); Swedish International Development Cooperation Agency (SIDA); the Swiss Agency for Development and Cooperation (SDC); and the Kenyan Government.https://www.mdpi.com/journal/pathogensam2022Zoology and Entomolog
Epidemiology of tick-borne pathogens of cattle and tick control practices in coastal Kenya
No full textTick-borne diseases (TBD) are a major constraint to livestock health and productivity in sub-Saharan Africa. Nonetheless, there are relatively few robust epidemiologic studies documenting TBD and its management in different endemic settings in Kenya. Therefore, a cross-sectional study using multi-stage cluster sampling was undertaken to characterize the epidemiology of TBD and management factors among zebu cattle reared under an extensive system in coastal Kenya. Blood samples from 1486 cattle from 160 herds in 14 villages were screened for the presence of tick-borne bacterial and protozoan pathogens using PCR with high-resolution melting analysis and sequencing. Standardized questionnaires were used to collect data on herd structure and herd management practices, and a mixed-effect logistic regression model to identify risk factors for tick-borne pathogens (TBPs). The application of chemical acaricide was the primary method for tick control (96.3%, 154/160), with the amidine group (mainly Triatix®, amitraz) being the most frequently used acaricides. Respondents identified East Coast fever as the most important disease and Butalex® (buparvaquone) was the most commonly administered drug in response to perceived TBD in cattle. The overall animal- and herd-level prevalence for TBPs were 24.2% (95% confidence interval (CI): 22.0–26.4%) and 75.6% (95% CI: 68.2–82.1%), respectively. Cattle were infected with Anaplasma marginale (10.9%, 95% CI: 9.4–12.6), Theileria parva (9.0%, 95% CI: 7.5–10.5), Anaplasma platys (2.6%, 95% CI: 1.9–3.6), Theileria velifera (1.1%, 95% CI: 0.7–1.8), Babesia bigemina (0.5%, 95% CI: 0.2–1.0), and Anaplasma sp. (0.1%, 95% CI: 0.0–0.4). Moreover, 21 cattle (1.4%) were co-infected with two TBPs. None of the assessed potential risk factors for the occurrence of either A. marginale or T. parva in cattle were statistically significant. The intra-herd correlation coefficients (lCCs) computed in this study were 0.29 (A. marginale) and 0.14 (T. parva). This study provides updated molecular-based information on the epidemiological status of TBPs of cattle and herd management practices in coastal Kenya. This information can be used in designing cost-effective control strategies for combating these TBD in the region
Proteome Profiling of Mycobacterium tuberculosis Cells Exposed to Nitrosative Stress
No full textReactive nitrogen species (RNS) are secreted by human cells in response to infection by Mycobacterium tuberculosis (Mtb). Although RNS can kill Mtb under some circumstances, Mtb can adapt and survive in the presence of RNS by a process that involves modulation of gene expression. Previous studies focused primarily on stress-related changes in the Mtb transcriptome. This study unveils changes in the Mtb proteome in response to a sub-lethal dose of nitric oxide (NO) over several hours of exposure. Proteins were identified using liquid chromatography coupled with electrospray ionization mass spectrometry (LC–MS/MS). A total of 2911 Mtb proteins were identified, of which 581 were differentially abundant (DA) after exposure to NO in at least one of the four time points (30 min, 2 h, 6 h, and 20 h). The proteomic response to NO was marked by two phases, with few DA proteins in the early phase and a multitude of DA proteins in the later phase. The efflux pump Rv1687 stood out as being the only protein more abundant at all the time points and might play a role in the early protection of Mtb against nitrosative stress. These changes appeared to be compensatory in nature, contributing to iron homeostasis, energy metabolism, and other stress responses. This study thereby provides new insights into the response of Mtb to NO at the level of proteomics
Anaplasma and Theileria Pathogens in Cattle of Lambwe Valley, Kenya: A Case for Pro-Active Surveillance in the Wildlife–Livestock Interface
No full textTick-borne pathogens (TBPs) are major constraints to livestock production and a threat to public health in Africa. This cross-sectional study investigated the risk of infection with TBPs in cattle of Lambwe Valley, Kenya. Blood samples of 680 zebu cattle from 95 herds in six geospatial clusters within 5 km of Ruma National Park were screened for bacterial and protozoan TBPs by high-resolution melting analysis and sequencing of PCR products. We detected Anaplasma bovis (17.4%), Anaplasma platys (16.9%), Anaplasma marginale (0.6%), Theileria velifera (40%), and Theileria mutans (25.7%), as well as an Anaplasma sp. (11.6%) that matched recently reported Anaplasma sp. sequences from Ethiopia. Babesia, Rickettsia, and Ehrlichia spp. were not detected. The animal and herd-level prevalences for TBPs were 78.5% (95% confidence intervals (CI): 75.3, 81.5) and 95.8% (95% CI: 91.8, 99.8), respectively. About 31.6% of cattle were co-infected with 13 combinations of TBPs. The prevalence of TBPs differed between clusters and age, but the risk of infection was not associated with sex, herd size, or the distance of homesteads from Ruma. This study adds insight into the epidemiology of TBPs around Ruma and highlights the need for proactive surveillance of TBPs in livestock–wildlife interfaces
Molecular characterization of Trypanosoma vivax in tsetse flies confirms the presence of the virulent Tvv4 genotype in Kenya : potential implications for the control of trypanosomiasis in Shimba Hills
No full textTrypanosoma vivax is a vector-borne protozoan parasite of livestock endemic to Africa and South America. To
date, fifteen genotypes of the parasite have been described in vertebrate and insect hosts in East Africa. However,
information regarding T. vivax diversity remains limited in many endemic countries in the sub-region, including
Kenya. Such information could deepen insight into the local epidemiology of animal trypanosomiasis in Shimba
Hills, a wildlife area in southeast Kenya where T. vivax is endemic and infects livestock. We employed two-gene
conventional-PCR-sequencing and phylogenetic analysis to characterize T. vivax genotypes in tsetse flies
collected between November 2018 and September 2019 in the wildlife-livestock interface of the Shimba Hills
National Reserve. Phylogenetic analysis of Internal Transcribed Spacer-1 (ITS-1) sequences of T. vivax isolates
confirmed the presence of two T. vivax genotypes in Shimba Hills of which >80% of T. vivax isolates from tsetse
flies clustered within the virulent Tvv4-genotype clade. Tsetse infections with the Tvv4 genotype were also
confirmed based on 18S rRNA gene sequencing. Expanded gene characterization identified three closely related
haplotypes within the Tvv4-clade. The Tvv4-isolates were detected in male and female Glossina pallidipes tsetse
flies, most of which were collected from grasslands and within two kilometres of the Shimba Hills National
Reserve boundary. Considering that T. vivax is the most common trypanosome in the Shimba Hills area and
causes severe clinical conditions in livestock, the Tvv4 genotype reported here for the first time in Kenya contributes
to our understanding of these pathologies. The effectiveness of trypanocidal drugs in the management of
Tvv4 is presently not clearly understood. Therefore, the parasite management in Shimba Hills should focus on
vector control to reduce the density of G. pallidipes, especially in grasslands near the wildlife protectorate.A German Academic Exchange Service (DAAD) in-region postgraduate scholarship; the BioVision Foundation Switzerland; European Union’s Integrated Biological Control Applied Research Programme – tsetse repellent component (EUIBCARP tsetse) awarded to icipe; UK’s Department for International Development (DFID); Swedish International Development Cooperation Agency (SIDA); the Swiss Agency for Development and Cooperation (SDC); and the Kenyan Government.http://www.elsevier.com/locate/meegidam2022Zoology and Entomolog
Biological control of ticks in domestic environments: Modeling the potential impact of entomopathogenic fungi on the transmission of East Coast fever in cattle
No full textBiological control of ticks using entomopathogenic fungi (EPF) is a highly desired alternative to chemical acaricides for the control of tick-borne pathogens. For Metarhizium anisopliae isolate ICIPE 7, one of these EPFs, efficacy against multiple tick species has been demonstrated in laboratory and field settings. However, we currently have little quantitative understanding of how EPFs can impact transmission. We developed a deterministic model of tick–host–pathogen interactions to explore how the effects of EPF on Rhipicephalus appendiculatus ticks may impact the transmission dynamics of East Coast fever (ECF) in cattle populations. We parameterized the multi-faceted effects of EPFs on tick dynamics using experimental data on Tickoff® biopesticide (a novel formulation of M. anisopliae ICIPE 7) and related EPFs. The epidemiological impact of EPF was evaluated across a range of product profiles and implementation strategies. Model results indicate that, for the explored product profiles, EPF derives most of its epidemiological impact through the delayed mortality effect. This EPF-induced mortality could not only reduce the onward Theileria parva transmission to cattle (both treated and untreated) but could also cause a reduction in the tick-to-host ratio and thus cattle exposure to ticks. The effects of EPF on reproduction fitness and engorgement of ticks elicit negligible impact. High levels of population coverage and treatment frequency are needed to reduce the tick population size and reach meaningful epidemiological impact in cattle populations. Additionally, increasing the persistence time of fungal conidia on cattle skin – through technological improvements to the EPF formulation–can substantially reduce acute infections when combined with appreciable population coverage levels, treatment frequency, and efficient spraying techniques. Our model analysis provides insights into the potential impact of EPF when deployed at a population level, and lends support to further research and development of this biological tick control tool
An investigation of the endocrine disrupting potential of enniatin B using <i>in vitro </i>bioassays
No full textEvidence that some of the fungal metabolites present in food and feed may act as potential endocrine disruptors is increasing. Enniatin B (ENN B) is among the emerging Fusarium mycotoxins known to contaminate cereals. In this study, the H295R and neonatal porcine Leydig cell (LC) models, and reporter gene assays (RGAs) have been used to investigate the endocrine disrupting activity of ENN B. Aspects of cell viability, cell cycle distribution, hormone production as well as the expression of key steroidogenic genes were assessed using the H295R cell model. Cell viability and hormone production levels were determined in the LC model, while cell viability and steroid hormone nuclear receptor transcriptional activity were measured using the RGAs. ENN B (0.01–100 μM) was cytotoxic in the H295R and LC models used; following 48 h incubation with 100 μM. Flow cytometry analysis showed that ENN B exposure (0.1–25 μM) led to an increased proportion of cells in the S phase at higher ENN B doses (>10 μM) while cells at G0/G1 phase were reduced. At the receptor level, ENN B (0.00156–15.6 μM) did not appear to induce any specific (ant) agonistic responses in reporter gene assays (RGAs), however cell viability was affected at 15.6 μM. Measurement of hormone levels in H295R cells revealed that the production of progesterone, testosterone and cortisol in exposed cells were reduced, but the level of estradiol was not significantly affected. There was a general reduction of estradiol and testosterone levels in exposed LC. Only the highest dose (100 μM) used had a significant effect, suggesting the observed inhibitory effect is more likely associated with the cytotoxic effect observed at this dose. Gene transcription analysis in H295R cells showed that twelve of the sixteen genes were significantly modulated (p < 0.05) by ENN B (10 μM) compared to the control. Genes HMGR, StAR, CYP11A, 3βHSD2 and CYP17 were downregulated, whereas the expression of CYP1A1, NR0B1, MC2R, CYP21, CYP11B1, CYP11B2 and CYP19 were upregulated. The reduction of hormones and modulation of genes at the lower dose (10 μM) in the H295R cells suggests that adrenal endocrine toxicity is an important potential hazard
A randomized controlled trial of Tickoff® (Metarhizium anisopliae ICIPE 7) for control of tick infestations and transmission of tick-borne infections in extensively grazed zebu cattle in coastal Kenya
No full textThe entomopathogenic fungus Metarhizium anisopliae isolate ICIPE 7 is being developed as an eco-friendly alternative to chemical acaricides in managing natural tick infestation on livestock. Its impact on tick infestation and tick-borne infections in cattle under natural conditions are yet unclear. We conducted a randomized controlled field trial to assess the safety and effects of Tickoff® (a formulation of M. anisopliae isolate ICIPE 7) and the chemical acaricide Triatix® on tick infestation and incidence of Anaplasma marginale and Theileria parva in extensively grazed zebu cattle in coastal Kenya. A total of 217 eligible herds comprising 1459 intent-to-treat zebu cattle were enrolled from 12 villages. The herds were randomly assigned in a 1:1:1 ratio to Tickoff®, Triatix®, or Tickoff® excipients. Tick counts, treatment administrations, and adverse events were registered every two weeks for seven months. The mortality of ticks collected from treated cattle was monitored in vitro. Infections with A. marginale and T. parva were monitored every two months. No adverse events were reported in either treatment group. Tickoff® did not significantly affect tick infestation (p = 0.869) or infection incidence (p > 0.05) compared to excipients. Triatix® significantly reduced tick infestation (p < 0.001) and incidence of T. parva (p = 0.042), but not A. marginale (p = 0.509) compared to the reference Tickoff®. In ticks that were removed from cattle, Tickoff® demonstrated significant pathogenicity in vitro relative to excipients (hazard ratio: 8.50, 95 % CI: 4.67–15.47). Fungus growth and sporulation were also observed on tick cadavers from Tickoff®, but not from excipients. While Tickoff® did not impact tick counts, its delayed, but significant effect on tick mortality may hinder onward pathogen transmission and give rise to indirect (i.e., to untreated animals) epidemiological effects, that were not picked up with this study design. Additionally, adverse environmental conditions resulted in low tick abundance and pathogen circulation towards the end of the study period, reducing the power of the study. This work re-emphasizes the challenges of randomized controlled field trials and the complexity of assessing the impact of vector control products on both direct and indirect impacts on pathogen transmission
