Goat Serum Research Articles

Development of ELISA kit for serological diagnosis of Q fever

In this study, it was aimed to develop an ELISA diagnostic kit that can be used in the serological investigation of Q fever disease caused by C. burnetii. For this purpose, bovine, sheep, goat and buffalo sera were tested with a commercial ELISA kit and home-made ELISA kit developed using two different conjugates. Of the 92 buffaloes blood serum samples tested, 22 (23.9%) were found positive by commercial ELISA kit, 7 (7.6%) and 29 (31.5%) were positive by protein-A conjugated ELISA and 29 anti-bovine IgG conjugated ELISA, respectively. In addition, Q fever positive and negative cattle, sheep, and goat blood sera that were found to be positive and negative for Q fever in a previous study conducted on animals with abortion history were also re-examined in our laboratory. In conclusion, within the scope of the project, an alternative homemade ELISA kit was developed for the serological diagnosis of Q fever and serum samples of the target species of buffalo, cattle, sheep, and goats were tested. Thus, the home-made ELISA kit, which can be used in research aimed at detecting seropositivity, especially in buffalo sera, was made ready for use. It was concluded that the data obtained would contribute to wider ranging epidemiological studies. The epidemiology of the disease can be revealed in detail in the light of research to be carried out with more comprehensive projects in our region.

Computational study to find the goat serum albumin (GSA) binding site as A-esterase

A-esterases are a classical term applied to enzymatic activity of the proteins by a mechanism not involving intermediate covalent phosphorylation, but requiring a divalent cation cofactor. Recently, a copper-dependent A-esterase activity has been identified in goat serum albumin (GSA) on the organophosphorus insecticide trichloronate. This hydrolysis was identified ex vivo with spectrophotometry and chromatography techniques. Albumin mechanism of action and catalytic site as Cu2+-dependent A-esterase are still unknown. Therefore, to know the copper bind to albumin is relevant. N-terminal sequence has been reported as the high affinity site for this cation, due to the histidine in position 3. The aim of this work in silico is to explore how occurs this metallic binding and active the esterase catalytic function. The GSA crystallized structure (PDB: 5ORI) was chosen for molecular docking and dynamics. A site-directed docking, for N-terminal site and a blind docking was done with trichloronate as ligand. Root-mean-square deviation and frequency plot was calculated to find the most frequent predicted structure and visualize the amino acids involved in binding site. The affinity energy in the blind docking (−5.80 kcal/mol) is almost twice lower than site-directed docking (−3.81 kcal/mol) and N-terminal amino acids do not appear in the most repeated structure binding site, suggesting that the protein has a site with higher affinity to the trichloronate ligand. His145 could be involved in the binding site as has been reported in previous studies.

Goats with low levels of AAV antibody may serve as candidates for large animal gene therapy

AAV vector-mediated gene therapy has been proposed as a feasible strategy for several eye diseases. However, AAV antibodies in the serum prior to treatment hinder the transduction efficiency and thus the therapeutic effect. Therefore, it is necessary to evaluate AAV antibodies in the serum before gene therapy. As large animals, goats are more closely related to humans than rodents and more economically available than nonhuman primates. Here, we first evaluated the AAV2 antibody serum level in rhesus monkeys before AAV injection. Then, we optimized a cell-based neutralizing antibody assay for detecting AAV antibodies in the serum of Saanen goats and evaluated the consistency of the cell-based neutralizing antibody assay and ELISA for goat serum antibody evaluation.The cell-based neutralizing antibody assay showed that the percentage of macaques with low antibody levels was 42.86%; however, there were no macaques with low antibody levels when the serum was evaluated by ELISA. The proportion of goats with low antibody levels was 56.67% according to the neutralizing antibody assay and 33. 33% according to the ELISA, and McNemar's test showed that the results of the two assays were not significantly different (P = 0.754), but that their consistency is poor (Kappa = 0.286, P = 0.114). Moreover, longitudinal evaluation of serum antibodies before and after intravitreal injection of AAV2 in goats revealed that the level of AAV antibodies increased and transduction inhibition subsequently increased, as reported in humans, indicating that transduction inhibition should be taken into account at different stages of gene therapy. In summary, starting with an evaluation of monkey serum antibodies, we optimized a detection method of goat serum antibodies, providing an alternative large animal model for gene therapy, and our serum antibody measurement method may be applied to other large animals.

Affinity-based electrochemical biosensor with antifouling properties for detection of lysophosphatidic acid, a promising early-stage ovarian cancer biomarker

Electrochemical techniques are considered to be highly sensitive, capable of fast response and can be easily miniaturized, properties which can aid with regard to the fabrication of compact point-of-care medical devices; however, the main challenge in developing such a tool is overcoming a ubiquitous, problematic phenomenon known as non-specific adsorption (NSA). NSA is due to the fouling of non-target molecules in the blood on the recognition surface of the device. To overcome NSA, we have developed an affinity-based electrochemical biosensor using medical-grade stainless steel electrodes and following a unique and novel strategy using silane-based interfacial chemistry to detect lysophosphatidic acid (LPA), a highly promising biomarker, which was found to be elevated in 90 % of stage I OC patients and gradually increases as the disease progresses to later stages. The biorecognition surface was developed using the affinity-based gelsolin-actin system, which was previously investigated by our group to detect LPA using fluorescence spectroscopy. We demonstrate the capability of this label-free biosensor to detect LPA in goat serum with a detection limit of 0.7 µM as a proof-of-concept for the early diagnosis of ovarian cancer.

APN Expression in Serum and Corpus Luteum: Regulation of Luteal Steroidogenesis Is Possibly Dependent on the AdipoR2/AMPK Pathway in Goats

Adiponectin (APN) is an essential adipokine for a variety of reproductive processes. To investigate the role of APN in goat corpora lutea (CLs), CLs and sera from different luteal phases were collected for analysis. The results showed that the APN structure and content had no significant divergence in different luteal phases both in CLs and sera; however, high molecular weight APN was dominant in serum, while low molecular weight APN was more present in CLs. The luteal expression of both AdipoR1/2 and T-cadherin (T-Ca) increased on D11 and 17. APN and its receptors (AdipoR1/2 and T-Ca) were mainly expressed in goat luteal steroidogenic cells. The steroidogenesis and APN structure in pregnant CLs had a similar model as in the mid-cycle CLs. To further explore the effects and mechanisms of APN in CLs, steroidogenic cells from pregnant CLs were isolated to detect the AMPK-mediated pathway by the activation of APN (AdipoRon) and knockdown of APN receptors. The results revealed that P-AMPK in goat luteal cells increased after incubation with APN (1 μg/mL) or AdipoRon (25 μM) for 1 h, and progesterone (P4) and steroidogenic proteins levels (STAR/CYP11A1/HSD3B) decreased after 24 h. APN did not affect the steroidogenic protein expression when cells were pretreated with Compound C or SiAMPK. APN increased P-AMPK and reduced the CYP11A1 expression and P4 levels when cells were pretreated with SiAdipoR1 or SiT-Ca, while APN failed to affect P-AMPK, the CYP11A1 expression or the P4 levels when pretreated with SiAdipoR2. Therefore, the different structural forms of APN in CLs and sera may possess distinct functions; APN might regulate luteal steroidogenesis through AdipoR2 which is most likely dependent on AMPK.

Molecular characterization and immunological properties of Echinococcus granulosus sensu stricto (G1) ADK1 and ADK8.

Adenylate kinases (ADKs) are one of the important enzymes regulating adenosine triphosphate (ATP) metabolism in Echinococcus granulosus sensu lato. The objective of the present study was to explore the molecular characteristics and immunological properties of E. granulosus sensu stricto (G1) adenylate kinase 1 (EgADK1) and adenylate kinase 8 (EgADK8). EgADK1 and EgADK8 were cloned and expressed, and the molecular characteristics of EgADK1 and EgADK8 were analyzed through different bioinformatics tools. Western blotting was used to examine the reactogenicity of recombinant adenylate kinase 1 (rEgADK1) and recombinant adenylate kinase 8 (rEgADK8) and to evaluate their diagnostic value. The expression profiles of EgADK1 and EgADK8 in 18-day-old strobilated worms and protoscoleces were analyzed by quantitative real-time PCR, and their distribution in 18-day-old strobilated worms, the germinal layer, and protoscoleces was determined by immunofluorescence localization. EgADK1 and EgADK8 were successfully cloned and expressed. Bioinformatics analysis predicted that EgADK1 and EgADK8 have multiple phosphorylation sites and B-cell epitopes. Compared with EgADK8, EgADK1 and other parasite ADKs have higher sequence similarity. In addition, both cystic echinococcosis (CE)-positive sheep sera and Cysticercustenuicollis-infected goat sera could recognize rEgADK1 and rEgADK8. EgADK1 and EgADK8 were localized in protoscoleces, the germinal layer, and 18-day-old strobilated worms. EgADK1 and EgADK8 showed no significant difference in their transcription level in 18-day-old strobilated worms and protoscoleces, suggesting that EgADK1 and EgADK8 may play an important role in the growth and development of E. granulosus sensu lato. Since EgADK1 and EgADK8 can be recognized by other parasite-positive sera, they are not suitable as candidate antigens for the diagnosis of CE.

Electrochemical Dopamine Detection Using Electrochemically Reduced Graphene Oxide/Silver/Metal Organic Framework Composites

AbstractA modified screen‐printed electrode based on zeolite imidazole framework‐8 (ZIF‐8), silver nanoparticles (Ag NPs) and electrochemically reduced graphene oxide (ErGO) were developed to detect dopamine. The morphology and electrochemical performance were investigated, and the results showed that ErGO/Ag‐ZIF‐8 composites were successfully synthesized without destruction of the ZIF‐8 structure. In addition, the peak current of dopamine (DA) oxidation increased linearly in the range of 0.1–200 μM, and the detection limit was 0.03 μM (S/N=3). Furthermore, the sensor showed good reproducibility and selectivity, and was successfully applied to the detection of DA in goat serum with a satisfactory recovery range.

Effect of N- carbamylglutamate and feeding level on amino acids and biochemical parameters in goats

The effect of feed additive N- carbamylglutamate (NCG) with different feeding levels on some biochemical parameters and amino acids in the serum of goats was investigated. Thirty male goats aged 10-11 months were distributed into three levels of concentrate 70%, 50%, and 30% with or without N- carbamylglutamate (NCG) in a 2×3 factorial experiment/ Completely Randomized Design (CRD). After 77 days of individual feeding, jugular blood was sampled before morning feeding. The results showed that additive NCG with high concentrate (70%) led to a decrease (P<0.01) in total blood protein, triglycerides, and globulins, and a decrease (P<0.05) for creatinine, uric acid, and VLDL. Blood magnesium increased (P<0.01) with NCG of low concentrate (30%) treatment, while phosphor increased with NCG of 50% concentrate treatment. Calcium increased with all NCG treatments. The results of adding N- carbamylglutamate with high roughage led to a negative effect on blood amino acids. In conclusion, increasing roughage feeding without NCG is the best for blood homeostasis, and don't use N- carbamylglutamate without non-protein nitrogen additives.
 Keywords: Amino acids, blood traits, N-carbamylglutamate, feeding level.

Application of UPLC-MS/MS Method for Analysis of Apigenin, Apigenin 7-Glucoside and Chlorogenic Acid in Goat Serum

UPLC-MS/MS method was developed for determination of bioactive compounds including chlorogenic acid (CA), apigenin (AP) and apigenin 7-glucoside (AP7G) in goat serum. The analytes were separated on Kinetex C18 analytical column and a mobile phase consisting 0.1% formic acid in water and methanol with gradient elution. Detection of analytes was performed using a triple quadrupole mass spectrometer with electrospray ionization in positive mode. The lower limit of quantitation was 0.5 ng mL–1 for AP and AP7G and 2.5 ng mL–1 for CA. The inter- and intra-day precision of the method expressed as relative standard deviation was in the range of 1.22–14.5%. The accuracy of the method expressed as the relative error was in the range of 0.147–14.6%. The extraction recoveries of the compounds were 76.2–89.5%. The validated method was successfully applied for analysis of the compounds in the serum of sixty goats, half of which were fed with fodder with the addition of an herbal mixture. The higher concentration values of all analyzed compounds were reported in the serum of the animals fed with herbs compared to the control group receiving standard fodder.

A Systematic Review and Meta-Analysis on Hepatitis E Virus Detection in Farmed Ruminants.

Swine are widely recognized as the main reservoir of zoonotic HEV; however, a growing body of data on the HEV prevalence in farmed ruminants of different species also points to a potential route for HEV transmission through ruminants and ruminant products and by-products. Definite information on the zoonotic potential of ruminants is still absent or unclear, compelling the necessity for increasing knowledge on this. The aim of the current study was to analyze the state-of-the-art in this research topic and provide a summary of HEV detection and characterization in farmed ruminants. A total of 1567 papers were retrieved from four search databases that resulted in 35 eligible papers after application of exclusion/inclusion criteria. Studies on HEV in farmed ruminants were mainly based on the detection of HEV RNA and were reported in Africa (n = 1), America (n = 3), Asia (n = 18) and Europe (n = 13), and focused on a variety of ruminants species, namely cow, goat, sheep, deer, buffalo and yak. The overall pooled prevalence of HEV was 0.02% (0.01-0.03, 95% CI). The subgroup pooled prevalence of HEV RNA was 0.01% (0.00-0.02, 95% CI) in cow milk, stool, serum, liver, intestinal, bile, blood, spleen and rectal swab samples; 0.09% (0.02-0.18, 95% CI) in goat serum, bile, stool, milk, liver, rectal swab and blood samples; 0.01% (0.00-0.04, 95% CI) in sheep stool, serum, milk, blood and liver samples. Most of the HEV genotypes found in farmed ruminants belonged to the zoonotic HEV-3 (subtypes 3a, 3c) and HEV-4 (subtype 4d, 4h), with Rocahepevirus also found. The wide HEV circulation observed in different farmed ruminants raises concerns for the possibility of HEV transmission through products from infected ruminants and alerts for the potential zoonotic route for HEV in ruminant products, such as meat and dairy products. Also, contact exposure to infected farmed animals could be a risk factor. Further research should be conducted in order to understand the circulation of HEV in these animals and its zoonotic potential, as there is currently a lack of data on this topic.

Highly Sensitive and Selective Dopamine Determination in Real Samples Using Au Nanoparticles Decorated Marimo-like Graphene Microbead-Based Electrochemical Sensors

A sensitive and selective electrochemical dopamine (DA) sensor has been developed using gold nanoparticles decorated marimo-like graphene (Au NP/MG) as a modifier of the glassy carbon electrode (GCE). Marimo-like graphene (MG) was prepared by partial exfoliation on the mesocarbon microbeads (MCMB) through molten KOH intercalation. Characterization via transmission electron microscopy confirmed that the surface of MG is composed of multi-layer graphene nanowalls. The graphene nanowalls structure of MG provided abundant surface area and electroactive sites. Electrochemical properties of Au NP/MG/GCE electrode were investigated by cyclic voltammetry and differential pulse voltammetry techniques. The electrode exhibited high electrochemical activity towards DA oxidation. The oxidation peak current increased linearly in proportion to the DA concentration in a range from 0.02 to 10 μM with a detection limit of 0.016 μM. The detection selectivity was carried out with the presence of 20 μM uric acid in goat serum real samples. This study demonstrated a promising method to fabricate DA sensor-based on MCMB derivatives as electrochemical modifiers.

Construction of a novel “self-regenerative” electrochemical biosensor based on metal–organic frameworks and its application to the detection of Mycoplasma ovine pneumonia

This study aimed to prepare a novel “self-regenerative” electrochemical biosensor by successively modifying gold nanoparticles, four-arm polyethylene glycol-NH2, and NH2-MIL-53 (Al) (MOF) on the glassy carbon electrode interface. A hairpin G-triplex-mediated DNA (G3 probe) as a part of the mycoplasma ovine pneumonia (MO) gene was loosely adsorbed to MOF. Based on the mechanism of hybridization induction, the G3 probe could effectively detach from the MOF only after introducing the target DNA. Subsequently, its guanine-rich nucleic acid sequences were exposed to solution of methylene blue. As a result, the diffusion current of the sensor system showed a sharp decline. The developed biosensor showed excellent selectivity, and the concentration of target DNA exhibited a good correlation in the range 10–10 to 10–6 M with a detection limit of 1.00 pM (S/N = 3), even in 10% goat serum. Most interestingly, this biosensor interface automatically started the regeneration program. Moreover, regeneration could be effectively achieved at least seven times, and the recovery rate of the electrode interface and sensing efficiency was up to 90%. Additionally, this platform could be used for other clinical assays in various systems by simply changing the DNA sequence of the probe.

P-072 Comparative study of indirect ELISA and AGID for serological detection of small ruminant lentiviruses in goats sera

P-072 Comparative study of indirect ELISA and AGID for serological detection of small ruminant lentiviruses in goats sera

High-performance enzyme-free glucose and hydrogen peroxide sensors based on bimetallic AuCu nanoparticles coupled with multi-walled carbon nanotubes

Interest in designing and manufacturing non-enzymatic biosensors is growing which stimulates the increased research efforts to explore innovative nanocomposites with highly catalytic activity. Here, the development of high-performance non-enzymatic glucose and hydrogen peroxide (H2O2) sensors based on bimetallic AuCu nanoparticles coupled with multi-walled carbon nanotubes (AuCu@MWCNTs) was achieved in this work. They were investigated by the cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and chronoamperometry (CA) methods separately. Benefiting from the synergistic effects of AuCu@MWCNTs, the proposed biosensors exhibited excellent performance such as low detection limits (1.3 nM for glucose, 0.1 nM for H2O2), broad linear ranges (8 nM-60 mM for glucose, 1 nM-4 mM for H2O2) and high sensitivities (362.3 μA·mM−1·cm−2 for glucose, 1335 μA·mM−1·cm−2 for H2O2). Besides, the AuCu@MWCNTs based electrodes provided excellent stability, repeatability and reproducibility. Based on the outstanding performance, they exhibited good reliability and accuracy in the detection of glucose in goat serum samples and real-time monitoring of H2O2 released by living cells. The present work suggests that the fabricated biosensor holds great potential for non-enzymatic glucose determination and real-time quantitative detection of H2O2 in biomedical applications.

Feeding Aquilaria sinensis Leaves Modulates Lipid Metabolism and Improves the Meat Quality of Goats.

Aquilaria (A.) sinensis is a medicinal plant widely grown in tropical South China. Given the abundant pruning waste of its leaves, the use of A. sinensis leaves is valuable. In this study, goats were fed a diet containing 20% A. sinensis leaves. Compared with the basal diet, feeding A. sinensis leaves to goats did not affect growth performance but considerably reduced the feeding cost. Strikingly, feeding A. sinensis leaves resulted in a significant decrease in the blood cholesterol levels (2.11 vs. 1.49 mmol/L, p = 0.01) along with a significant increase in the high-density lipoprotein levels (1.42 vs. 1.82 mmol/L, p = 0.01). There was also a tendency to lower the content of low-density lipoprotein levels in goats (0.78 vs. 0.45 mmol/L, p = 0.09). Furthermore, metabolomics analysis demonstrated that the reduction in cholesterol levels occurred in both the serum (0.387-fold change) and muscle (0.382-fold change) of goats during A. sinensis leaf feeding. The metabolic responses to feeding A. sinensis leaves suggest that the activation of lipolysis metabolism might happen in goats. These observed changes would be conducive to improving animal health and meat quality, ultimately benefiting human health.

Sero-epidemiology of brucellosis in small ruminants on rangeland in northern Mexico

The seroprevalence of Brucella antibodies in serum of goats and sheep reared on semi-arid pastoral systems of northeastern Mexico was assessed. Additionally, species, gender, and predominant breed were evaluated as risk factors for seropositivity to brucellosis. Data were from 375 flocks of goats or co-mingled goats and sheep. Serum samples from 11001 goats and 4741 sheep collected between 2016 and 2019 were analyzed with the brucellosis card test. The disease affected 12% of flocks. The overall prevalence of brucellosis seropositivity in goats and sheep were estimated at 2.1% (95% CI = 1.78-2.31) and 0.8% (95% CI = 0.58-1.10), respectively. Brucellosis seropositivity was two times more likely (p < 0.01) in goats than sheep. The risk of seropositivity to brucellosis for goats and sheep was 2.3 and 3.2 times higher (p < 0.01) in females than males. Toggenburg goats were more likely (p < 0.01) to be seropositive to brucellosis than all other breeds of goats, whereas criollo sheep were more likely to be seropositive to brucellosis compared to hair sheep composite breeds. The current study revealed that, despite the absence of Brucella vaccination, brucellosis is not widely distributed in the study area. Also of all animals screened, seropositivity to Brucella infection was highest in Toggenburg when compared to other dairy and meat breeds; Criollo sheep also presented the highest seropositivity to brucellosis compared to hair sheep composite breeds. Finally, females compared to males had increased odds of testing positive for brucellosis.

Effects of fermented feed of Pennisetum giganteum on growth performance, oxidative stress, immunity and gastrointestinal microflora of Boer goats under thermal stress.

This study was conducted to evaluate the effects of fermented feed of Pennisetum giganteum (P. giganteum) on growth performance, oxidative stress, immunity and gastrointestinal microflora of Boer goats under thermal stress. The study was conducted during 45 days using twenty 2 months Boer goats. The goats were randomly allocated into two groups: NPG (n = 10; normal P. giganteum) and FPG (n = 10; fermented feed of P. giganteum), and the ratio of concentrates to roughage was 3:2. Both groups of animals were kept in sheds and exposed to summer thermal stress from 10:00 h to 18:00 h (temperature and humidity index, THI > 78). At the end of the study, the animals were slaughtered and assessed for various characteristics. The findings from the study revealed that FPG-feeding significantly increased (p < 0.05) average daily gain (ADG, 48.18 g) and carcass weight (4.38 kg), while decreased (p < 0.01) average daily feed intake (ADFI, 0.74 kg/d; p < 0.01) and the feed:gain (F/G, 15.36) ratio. The CAT, GSH-Px activities and GSH in serum, liver and spleen, and the levels of IgA, IgG, IgM, IL-2, IL-4 and IL-1β in serum of FPG-fed goats were significantly higher (p < 0.05) than those of NPG-feeding goats. Further, we found that FPG feed is rich in nutrients with Lactobacillus (65.83%) and Weissella (17.80%). Results for gastrointestinal microbiota composition showed that FPG-feeding significantly enhanced the abundance of Lactobacillus and unidentified Clostridiales, and reduced Anaerovibrio and Methanobrevibacter. Meanwhile, Spearman's correlation analysis showed that these microbiotas were closely related to the improvement of oxidative stress and immune indexes of goats. These results demonstrated that FPG-feeding not only reduces oxidative stress and improves ROS clearance to enhance antioxidant defense system, but also improves gastrointestinal microbiota to enhance immune function by overcoming the adverse effects of heat stress, and further improve growth performance of goats.

Effect of N- carbamylglutamate on amino acids and blood biochemistry in young male goats

To study the effect of feed additive N- carbamylglutamate (NCG) with different levels of feeding on amino acids profile and some analytes in serum of goats, thirty male goats aged 7-8 months were distributed into three levels of concentrate 2%, 3%, 4% with or without NCG in a 2×3 factorial experiment/ Completely Randomized Design (CRD). After 72 days of individual feeding, the blood from jugular vein was obtained before feeding in the morning. Results showed that additive NCG with 3% concentrate led to an increase (P&lt;0.01) in total protein and albumin (P&lt;0.05), globulins increased (P&lt;0.01) with increasing concentrate without NCG. Creatinine decreased with increasing concentrate without NCG. Additive NCG led to a linear decrease in uric acid with increasing feed intake (P&lt;0.05) and a linear increase without NCG. Blood magnesium, zinc, and copper increased with NCG. No difference in calcium between treatments. The level of feeding without NCG didn't affect fat parameters, while LDL, HDL, and cholesterol increased with NCG and 4% concentrate. Additive N- carbamylglutamate affected negatively blood amino acids, especially with 4% concentrate. In conclusion, 3% concentrate was the best for blood homeostasis with or without NCG, and preferred to use NCG with urea.

Comparative Analysis of Tick-Borne Relapsing Fever Spirochaetes from Ethiopia and Nigeria.

Despite increasing reports of tick-borne diseases in Africa, remarkably, reports of tick-borne relapsing fever (TBRF) in Nigeria are lacking. Ornithodoros savignyi from Nigeria have been reported with the relapsing fever Candidatus Borrelia kalaharica. Conversely, in Ethiopia, the agent of relapsing fever is the louse-borne relapsing fever (LBRF) spirochaete Borrelia recurrentis with no TBRF reported to occur. A total of 389 Ornithodoros ticks, Ethiopia (N = 312) and Nigeria (N = 77), were sampled, together with 350 cattle, and 200 goat sera were collected from Nigeria. Samples were screened for Borrelia spp. by RT-PCR. Reactive samples were confirmed, then sequenced using flagellin B, 16S rRNA, and 16S-23S intergenic spacer region. The prevalence of Borrelia spp. in livestock was 3.8% (21/550) and 14% (3/21) after final molecular confirmation. Of 312 ticks from Ethiopia, 3.5% (11/312) were positive for Borrelia, with 36% (4/11) by conventional PCR. Sequencing revealed that the borreliae in soft ticks was C. B. kalaharica, whilst that found in animals was Borrelia theileri. Soft ticks were confirmed by sequencing 7% (22/312) and 12% (9/77) of the Ethiopian and Nigerian ticks, respectively. Phylogenetic analysis revealed that these were Ornithodoros savignyi. This is the first evidence of C. B. kalaharica in Ethiopia and demonstrates the co-existence of TBRF in a country endemic to LBRF. Important, this might cause a diagnostic challenge given that LBRF is predominantly diagnosed by microscopy, which cannot differentiate these two spirochaetes. Furthermore, we report B. theileri in ruminants in Nigeria, which may also be of veterinary and economic importance.

One-step synthesis of 2D@3D hollow Prussian blue analogue as a high-performance bifunctional electrochemical sensor.

Prussian blue analogues (PBAs) are a family of classic coordination polymers. They have been widely applied in various fields including electrochemical sensors. Cubic nanoparticle structure is their common morphology. It is still a great challenge to design a hollow and two-dimensional (2D) PBA material. Of course, it will be a significant surprise if a hollow cube and 2D sheet can be integrated into one material. In this work, we designed a simple one-step synthetic strategy and resolved the above difficulty, wherein a hollow cubic PBA covered by 2D ultrathin nanosheets was successfully constructed, namely hollow tremella-like PBA (HTPBA). Furthermore, Ni foam (NF) as a substrate was introduced to obtain a self-supporting HTPBA/NF-12 electrode. HTPBA/NF-12, as a bifunctional electrochemical sensor electrode, exhibited distinguished catalytic performance towards glucose and nitrite, including remarkable selectivity, reproducibility, sensitivity for glucose (21 410 μA mM-1 cm-2) and nitrite (1248 μA mM-1 cm-2), wide linear range of 2-1250 μM and 5-3380 μM, along with low detection limit of 0.056 μM and 0.38 μM, respectively. More importantly, HTPBA/NF-12 electrodes possessed good reusability and practicability even in goat serum. In this study, we developed a simple and effective strategy to fabricate 2D@3D PBA material with excellent electrocatalytic activity and provide a totally new viewpoint in the PBA sensing field.