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Abstract Background Innovations in cancer treatment have contributed to the improved survival rate of cancer patients. The cancer survival rates have been growing and nearly two third of those survivors have been exposed to clinical radiation during their treatment. The study of long-term radiation effects, especially secondary cancer induction, has become increasingly important. An accurate assessment of out-of-field/peripheral dose (PDs) is necessary to estimate the risk of second cancer after radiotherapy and the damage to the organs at risk surrounding the planning target volume. This study was designed to measure the PDs as a function of dose, distances, and depths from Telecobalt-60 (Co-60) beam in water phantom using thermoluminescent dosimeter-100 (TLD-100). Methods The PDs were measured for Co-60 beam at specified depths of 0 cm (surface), 5 cm, 10 cm, and 15 cm outside the radiation beam at distances of 5, 10, and 13 cm away from the radiation field edge using TLD-100 (G1 cards) as detectors. These calibrated cards were placed on the acrylic disc in circular tracks. The radiation dose of 2000 mGy of Co-60 beam was applied inside 10 × 10 cm2 field size at constant source to surface distance (SSD) of 80 cm. Results The results showed maximum and minimum PDs at surface and 5 cm depth respectively at all distances from the radiation field edge. Dose distributions out of the field edge with respect to distance were isotropic. The decrease in PDs at 5 cm depth was due to dominant forward scattering of Co-60 gamma rays. The increase in PDs beyond 5 cm depth was due to increase in the irradiated volume, increase in penumbra, increase in source to axis distance (SAD), and increase in field size due to inverse square factor. Conclusion It is concluded that the PDs depends upon depth and distance from the radiation field edge. All the measurements show PDs in the homogenous medium (water); therefore, it estimates absorbed dose to the organ at risk (OAR) adjacent to cancer tissues/planning target volume (PTV). It is suggested that PDs can be minimized by using the SAD technique, as this technique controls sources of scattered radiation like inverse square factor and effect of penumbra up-to some extent.

Haemonchus contortus poses a global challenge as a parasite affecting small ruminants, yet the problem of absence of an effective vaccine against H. contortus infection still exists. This investigation sought to appraise the immunological reaction induced by recombinant H. contortus excretory/secretory-24 (rHcES-24) in combination with complete Freund’s adjuvant (CFA) and bio-polymeric nanoparticles (NPs) within a murine model. In this study, rHcES-24 was encapsulated in poly(d, l-lactide-co-glycolide) (PLGA) and chitosan (CS) NPs, administered subcutaneously to mice. Researchers analyzed the NPs using scanning electron microscope (SEM) and assessed lymphocyte proliferation, specific antibodies, cytokines, T cell proliferation (CD3e+CD4+, CD3e+CD8a+), and phenotypic alteration in splenocytes (CD11c+CD83+, CD11c+CD86+) through flow cytometry to understand the immune response. The results demonstrated that the administration of nanovaccines (NVs) prompted immune responses towards Th1 pathway. This was indicated by notable enhancements in the production of specific antibodies, heightened cytokine levels, and a robust proliferation of lymphocytes observed in mice that received the NVs compared to control groups. Remarkably, mice vaccinated with the antigen-loaded NPs formulations exhibited considerably higher proportions of splenic dendritic cells (DCs) and T cells in comparison to those receiving the traditional adjuvant or the control groups. Incorporating HcES-24 protein into NPs effectively conferred immunity against H. contortus, paving the way for developing a targeted and commercial vaccine.

In this study, we analyze complex partial differential equations, specifically focusing on nonlinear fractional-order Inviscid Burger's equations. By employing the Adomian Decomposition Method, we develop analytical approximations in the form of convergent series to solve these equations with time and space fractional derivatives in the Caputo sense. Our approach provides explicit, efficient, and highly accurate solutions, which we verify against exact solutions, eliminating the need for closure approximations and perturbation theory. We present our findings through detailed tables and graphs, offering valuable insights into the behavior of the solutions. This work advances analytical techniques and establishes a foundation for new applications in various scientific and engineering fields.

Background: Haemonchus contortus (H. contortus), a nematode with global prevalence, poses a major threat to the gastrointestinal health of sheep and goats. In an effort to combat this parasite, a nanovaccine was created using a recombinant ADP-ribosylation factor 1 (ARF1) antigen encapsulated within poly lactic-co-glycolic acid (PLGA). This study aimed to assess the effectiveness of this nanovaccine in providing protection against H. contortus infection. Methods: Fifteen goats were randomly divided into three groups. The experimental group received two doses of the PLGA encapsulated rHcARF1 (rHcARF1-PLGA) nanovaccine on days 0 and 14. Fourteen days after the second immunization, both the experimental and positive control groups were challenged with 8000 infective larvae (L3) of H. contortus, while the negative control group remained unvaccinated and unchallenged. At the end of the experiment on the 63rd day, all animals were humanly euthanized. Results: The results showed that the experimental group had significantly higher levels of sera IgG, IgA, and IgE antibodies, as well as increased concentrations of cytokines, such as IL-4, IL-9, IL-17, and TGF-β, compared to the negative control group after immunization. Following the L3 challenge, the experimental group exhibited a 47.5% reduction in mean eggs per gram of feces (EPG) and a 55.7% reduction in worm burden as compared to the positive control group. Conclusions: These findings indicate that the nanovaccine expressing rHcARF1 offers significant protective efficacy against H. contortus infection in goats. The results also suggest the need for more precise optimization of the antigen dose or a reassessment of the vaccination regimen. Additionally, the small sample size limits the statistical rigor and the broader applicability of the findings.

Bacillus thuringiensis (B. thuringiensis) is considered one of the most important entomopathogenic microorganisms. It produces potent toxins against insects. Therefore, the present study investigates the bioacaricidal properties of B. thuringiensis on the Hyalomma tick species. Firstly, we identify Hyalomma ticks based on morphological screening and molecular characterization. The cytochrome C oxidase subunit I (COX1) gene was selected for the polymerase chain reaction (PCR) analysis, which resulted in the amplification of 656 bp. The amplified products were sequenced, and the isolated (COX1) gene of ticks was submitted to the gene bank of NCBI (Accession No. OR077934.1). The nucleotide sequences were retrieved from the NCBI data bank by BLASTn analysis, which confirmed that all obtained sequences belong to genus Hyalomma, and multiple alignments confirmed that the sequence of Hyalomma anatolicum Tandojam-isolate (HA-TJ) 100% aligned with Hyalomma analoticum KP792577.1, Hyalomma detritum KP792595.1, Hyalomma excavatum KX911989.1, and H. excavatum OQ449693.1. The generated phylogenetic tree confirmed that sequences of HA-TJ COX1 clustered with a single clad of H. analoticum, H. excavatum, and H. detritum. The acaricidal effect of B. thuringiensis toxins B. thuringiensis spore crystal mix (BtSCM) and B. thuringiensis crystal proteins (Btcps) was evaluated against larvae and adult life stages of Hyalomma ticks in vitro. We applied Btcps and BtSCM separately with different concentrations and calculated the mortality percentage. Adult mortality was estimated at the 8th, 10th, 12th, and 15th days posttreatment and larval mortality after 24 h. During treatment of the adult life stage, at first, ticks were immersed in different concentrations of Btcps and BtSCM for 5 min after the treatments, and the samples were transferred to sterile containers and placed in an incubator with 80% humidity at 23°C. Furthermore, Btcps produced the highest mortality on Day 15, 89±1.00% at a concentration of 3000 μg/mL, followed by the 12th, 10th, and 8th days produced 83±1.91%, 70±1.15%, and 61±1.00%, respectively. BtSCM produced mortality of 69±1.91% on Day 15 at a concentration of 3000 μg/mL, followed by the 12th, 10th, and 8th days at 57±2.51%, 37±1.91%, and 34±2.00%. The present study revealed that B. thuringiensis toxins produced a significant (p

Babesiosis is a protozoal disease affect livestock and pet animals such as cattle, buffaloes, sheep, goats, horses, donkeys, mules, dogs, and cats. It causes severe economic losses in livestock as well as in pet animals. A large number of dairy animals are imported in order to fulfill the demands of milk, milk, meat and its products. In addition, different pet animals are transported from Pakistan to various parts of the world, therefore, it is important to identify the current status and distribution of babesiosis throughout Pakistan in order to control the disease and draw attention for future research, diagnosis, treatment and control of this diseases. No work has been done on a complete review on up-to-date on blood protozoal disease burden in Pakistan. This article will provide about the complete background of babesiosis in ruminants, equines and pet animals, its current status, distribution, vectors in Pakistan and allopathic and ethnoveterinary treatments used against babesiosis. Babesiosis may be subclinical (apparently normal) and may be clinical with acute to chronic disease and sometimes fatal. Babesia is found and develops inside the erythrocytes (red blood cells). Clinically, it causes fever, fatigue, lethargy, pallor mucus membranes, malaise, cachexia, respiratory distress, jaundice, icterus, hemolytic anemia, hemoglobinuria, lymphadenopathy, chollangocytitis, hepatomegaly, and splenomegaly. Chemotherapy for babesiosis includes Imidocarb dipropionate, Diaminazine aceturate Atovaquone and Bupravaquone, Azithromycin, Quinuronium sulfate and Amicarbalidesio-thionate are most widely used. Supportive therapy includes multivitamins, fluid therapy, antipyretics intravenous fluids, and blood transfusions are used if necessary. In addition, there are certain ethnoveterinary (homeopathic) ingredients which having anti-babesial activity. As the resistance against these drugs is developing every day. New more specific long-lasting drugs should be developed for the treatment of Babesiosis. Further studies should be done on disease genome of different species of Babesia for vaccine development like malarial parasites.

This study presents a structure preserving nonstandard finite difference scheme to analyze a susceptible-infected-treatment-recovered (SITR) dynamical model of coronavirus 2019 (covid-19) with bimodal virus transmission in susceptible population. The underlying model incorporates the possible treatment measures as the emerging scenario of covid-19 vaccines. Keeping in view the fact that the real time data for covid-19 is updated at discrete time steps, we propose a new structure preserving numerical scheme for the proposed model. The proposed numerical scheme produces realistic solutions of the complex bi-modal SITR nonlinear model, converges unconditionally to steady states and reflects dynamical consistency with continuous sense of the model. The analysis of the model reveals that the model remains stable at the steady state points. The basic reproduction number Rcovid falls less than 1 when treatment rate is increased and disease will die out. On the other hand, it predicts that human population may face devastating effects of pandemic if the treatment measures are not strictly implemented.

This study proposes a Padé approximation based hybrid mesh free framework for numerical solutions of nonlinear partial differential equations. The proposed framework involves three novel aspects. Firstly, the conjunction of Padé approximation based residual functional and penalty function approach for handling boundary/Initial conditions is employed to design an equivalent optimization problem. Secondly, the notion of cooperative multi-simplex searches in the environment of Nelder-Mead Simplex (NMS) algorithm is introduced. Finally, a hybrid of multi-simplex search and Differential Evolution (DE) is proposed to efficiently cope with the modelled problem. The developed method is named as Padé Evolutionary Cooperative Multi-Simplex (PECMS) algorithm. The results of the developed PECMS scheme on illustrative examples are compared with three well-known methods, namely, Differential Transform Method (DTM), Adomian Decomposition Method (ADM) and the Variational Iteration Method (VIM). The comparisons demonstrate that the current approach possesses high accuracy when compared to the competing methods.

Economic load dispatch problems are most important in operation and management of the electric power systems which are formulated as optimization problems. Modern deterministic and stochastic optimization techniques are efficient in solving economic load dispatch problems without any limitation because of their capability to seek the global optimal solution. Economic load dispatch problems involve linear equality constraints which are difficult to handle along with discontinuous, non-differentiable and highly non-linear objective functions. The problem becomes challenging for optimization techniques, especially for deterministic methods. This study presents a new approach based on a hybrid algorithm consisting of Genetic algorithm and modified Hooke and Jeeves method to solve the economic load dispatch problems with equality constraints. The performance of proposed Algorithm is tested on five generating systems with valve-point effects. Test results of proposed technique are quite promising and effective, with good convergence property and generation cost to produce better quality solution compared with several state-of-the-art methods.

Abstract Background High concentrate (HC) diet-induced oxidative stress causes gut epithelial damages associated with apoptosis. Selenium (Se) being an integral component of glutathione peroxidase (GSH-Px) plays an important role in antioxidant defense system. Therefore, increasing dietary Se level would alleviate HC diet-induced injuries in gut mucosa. The present study investigated eighteen cross-bred goats, randomly divided into three groups (n = 6/group) fed either low concentrate (LC, roughage: concentrate ratio 65:35), high concentrate (HC, 35:65) or HC plus Se (HC-SY) diets for 10 weeks. Se was supplemented at the dose rate of 0.5 mg Se kg− 1 diet in the form of selenium yeast. The background Se level in HC and LC diets were 0.15 and 0.035 mg.kg− 1 diet, respectively. The Se at the dose of 0.115 mg.kg− 1 diet was added in LC diet to make its concentration equivalent to HC diet and with the supplementation of 0.5 mg Se kg− 1, the goats in group HC-SY received total Se by 0.65 mg.kg− 1 diet. Results The molar concentrations of individual and total short chain fatty acids (TSCFA) significantly increased (P

This study presents numerical work to investigate the Falkner–Skan flow of a bio-convective Casson fluid over a wedge using an Evolutionary Padé Approximation (EPA) scheme. The governing partial differential equations and boundary conditions of a Falkner–Skan flow model are transformed to a system of ordinary differential equations involving ten dimensionless parameters by using similarity transformations. In the proposed EPA framework, an equivalent constrained optimization problem is formed. The solution of the resulting optimization problem is analogous to the solution of the dimensionless system of ordinary differential equations. The solutions produced in this work, with respect to various combinations of the physical parameters, are found to be in good agreement with those reported in the previously published literature. The effects of a non-dimensional physical-parameter wedge, Casson fluid, fluid phase effective heat capacity, Brownian motion, thermophoresis, radiation, and magnetic field on velocity profile, temperature profile, fluid concentration profile, and the density of motile microorganisms are discussed and presented graphically. It is observed that the fluid velocity rises with a rise in the Casson fluid viscosity force parameter, and an increase in the Prandtl number causes a decrease in the heat transfer rate. Another significant observation is that the temperature and fluid concentration fields are greatly increased by an increase in the thermophoresis parameter. An increase in the Péclet number suppresses the microorganism density. Moreover, the increased values of the Prandtl number increase the local Nusslet number, whereas the skin friction is increased when an increase in the Prandtl number occurs.

Abstract Babesia spp. are tick-transmitted apicomplexan parasites that infect erythrocytes of a wide range of vertebrates including domestic animals, and can be associated with a considerable economic loss due to introduction of prophylactic measures, decreased production and rates of morbidity and mortality. An efficient detection assay needs to be set up for accurate diagnosis and prevention of disease. The present work is aimed at verifying the occurrence of Babesia bigemina infection in water buffaloes in the southeastern region of Pakistan by using both traditional blood smear and molecular techniques. A total of 100 blood samples were collected from tick-free and tick-infested animals (50 animals each group). The overall prevalence was recorded as 20% and 17% by using thin blood smear and PCR methods, respectively. Moreover, the analysis of infection in tick-infested and tick-free animals showed the infection rates of 36% and 4% by using blood smear method, and 30% and 4% with that of PCR method, respectively. Despite the higher prevalence rate recorded by blood smear method, which may be the result of non-specific identification of Babesia species, these results indicate that the PCR assay used in this study provides a useful tool for accurate diagnosis of the Babesia bigemina infection in bovines. Key words: Babesia bigemina, diagnosis, PCR, prevalence, blood smear technique

This work presents a novel evolutionary computation-based Padé approximation (EPA) scheme for constructing a closed-form approximate solution of a nonlinear dynamical model of Covid-19 disease with a crowding effect that is a growing trend in epidemiological modeling. In the proposed framework of the EPA scheme, the crowding effect-driven system is transformed to an equivalent nonlinear global optimization problem by assimilating Padé rational functions. The initial conditions, boundedness, and positivity of the solution are dealt with as problem constraints. Keeping in view the complexity of formulated optimization problem, a hybrid of differential evolution (DE) and a convergent variant of the Nelder-Mead Simplex algorithm is also proposed to obtain a reliable, optimal solution. The comparison of the EPA scheme results reveals that optimization results of all formulated optimization problems for the Covid-19 model with crowding effect are better than those of several modern metaheuristics. EPA-based solutions of the Covid-19 model with crowding effect are in good agreement with those of a well-practiced nonstandard finite difference (NSFD) scheme. The proposed EPA scheme is less sensitive to step lengths and converges to true equilibrium points unconditionally.

The Plateau-Bézier problem with shifted knots is to find the surface of minimal area amongst all the Bézier surfaces with shifted knots spanned by the admitted boundary. Instead of variational minimization of usual area functional, the quasi-minimal Bézier surface with shifted knots is obtained as the solution of variational minimization of Dirichlet functional that turns up as the sum of two integrals and the vanishing condition gives us the system of linear algebraic constraints on the control points. The coefficients of these control points bear symmetry for the pair of summation indices as well as for the pair of free indices. These linear constraints are then solved for unknown interior control points in terms of given boundary control points to get quasi-minimal Bézier surface with shifted knots. The functional gradient of the surface gives possible candidate functions as the minimizers of the aforementioned Dirichlet functional; when solved for unknown interior control points, it results in a surface of minimal area called quasi-minimal Bézier surface. In particular, it is implemented on a biquadratic Bézier surface by expressing the unknown control point P11 as the linear combination of the known control points in this case. This can be implemented to Bézier surfaces with shifted knots of higher degree, as well if desired.

Abstract This work proposes a novel mesh free evolutionary Padé approximation (EPA) framework for obtaining closed form numerical solution of a nonlinear dynamical continuous model of virus propagation in computer networks. The proposed computational architecture of EPA scheme assimilates a Padé approximation to transform the underlying nonlinear model to an equivalent optimization problem. Initial conditions, dynamical positivity and boundedness are dealt with as problem constraints and are handled through penalty function approach. Differential evolution is employed to obtain closed form numerical solution of the model by solving the developed optimization problem. The numerical results of EPA are compared with finite difference schemes like fourth order Runge–Kutta (RK-4), ODE45 and Euler methods. Contrary to these standard methods, the proposed EPA scheme is independent of the choice of step lengths and unconditionally converges to true steady state points. An error analysis based on residuals witnesses that the convergence speed of EPA is higher than a globally convergent non-standard finite difference (NSFD) scheme for smaller as well as larger time steps.

Abstract Background Arginine kinase (AK), an important member of phosphagen kinase family has been extensively studied in various vertebrates and invertebrates. Immunologically, AKs are important constituents of different body parts, involved in various biological and cellular functions, and considered as immune-modulator and effector for pro-inflammatory cytokines. However, immunoregulatory changes of host cells triggered by AK protein of Haemonchus contortus, a parasitic nematode of ruminants, are still unknown. The current study was focused on cloning and characterisation of Hc-AK, and its regulatory effects on cytokines level, cell migration, cell proliferation, nitric oxide production and apoptosis of goat peripheral blood mononuclear cells (PBMCs) were observed. Methods The full-length sequence of the Hc-AK gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sub-cloned into the prokaryotic expression vector pET-32a. The biochemical characteristics of recombinant protein Hc-AK, which was purified by affinity chromatography, were performed based on the enzymatic assay. Binding of rHc-AK with PBMCs was confirmed by immunofluorescence assay (IFA). Immunohistochemical analysis was used to detect localisation of Hc-AK within adult worms sections. The immunoregulatory effects of rHc-AK on cytokine secretions, cell proliferation, cell migration, nitric oxide production and apoptosis were determined by co-incubation of rHc-AK with goat PBMCs. Results The full-length ORF (1080 bp) of the Hc-AK gene was successfully cloned, and His-tagged AK protein was expressed in the Escherichia coli strain BL21. The recombinant protein of Hc-AK (rHc-AK) was about 58.5 kDa together with the fused vector protein of 18 kDa. The biochemical assay showed that the protein encoded by the Hc-ak exhibited enzymatic activity. Western blot analysis confirmed that the rHc-AK was recognised by the sera from rat (rat-antiHc-AK). The IFA results showed that rHc-AK could bind on the surface of goat PBMCs. Immunohistochemically, Hc-AK was localised at the inner and outer membrane as well as in the gut region of adult worms. The binding of rHc-AK to host cells increased the levels of IL-4, IL-10, IL-17, IFN-γ, nitric oxide (NO) production and cell apoptosis of goat PBMCs, whereas, TGF-β1 levels, cell proliferation and PBMCs migration were significantly decreased in a dose dependent manner. Conclusions Our findings suggested that rHc-AK is an important excretory and secretory (ES) protein involved in host immune responses and exhibit distinct immunomodulatory properties during interaction with goat PBMCs.

Abstract Background Aspartyl protease inhibitor (API) was thought to protect intestinal parasitic nematodes from their hostile proteolytic environment. Studies on Ostertagia ostertagi, Ascaris suum and Brugia malayi indicated that aspins might play roles in nematode infection. In a recent study, proteins differentially expressed between free-living third-stage larvae (L3) and activated L3 (xL3) of Haemonchus contortus were identified by 2D-DIGE. API was found downregulated in xL3 when compared with L3. However, there was no report about the functions of H. contortus API in the parasite-host interaction. In this study, the gene encoding API from H. contortus was cloned, expressed, and part of its biological characteristics were studied. Results A DNA fragment of 681 bp was amplified by RT-PCR. Ninety one percent of the amino acid sequence was similar with that for aspin from O. ostertagi. The recombinant API protein was fusion-expressed with a molecular weight of 48 × 103. Results of Western blot showed that the recombinant API could be recognized by serum from goat infected with H. contortus. It was found that API was localized exclusively in the subcutaneous tissue and epithelial cells of the gastrointestinal tract in adult H. contortus. qRT-PCR suggested that the API gene was differentially transcribed in different life-cycle stages, with the lowest level in female adults and the highest in free-living L3 larvae. Enzyme inhibition assay indicated that the recombinant API can inhibit the activity of pepsin significantly, and the optimal reaction pH and temperature were 4.0 and 37–50 °C respectively. In vitro study showed that the recombinant API could induce goat PBMCs to express IFN-γ, IL-4 and IL-10. Conclusions A new aspartyl protease inhibitor was cloned from H. contortus and its characteristics were studied for the first time. The results indicate that API may regulate the immune response of the host and play roles in the infection.

ADP-ribosylation factor 1 (HcARF1) is one of the Haemonchus contortus (H. contortus) excretory/secretory proteins involved in modulating the immune response of goat peripheral blood mononuclear cells (PBMC). Here, we evaluated the immunogenic potential of recombinant HcARF1 (rHcARF1) against H. contortus infection in Institute of Cancer Research (ICR) mice. Briefly, rHcARF1 was entrapped in poly (D, L-lactide-co-glycolide) (PLGA) and chitosan (CS) nanoparticles (NP) and injected into mice as a vaccine. Fifty-six ICR mice were assigned randomly into seven groups, with eight animals in each group, and they were vaccinated subcutaneously. At the end of the experiment (14th day), the blood and the spleen were collected from euthanized mice to detect lymphocyte proliferation, cytokine analysis, and the production of antigen-specific antibodies. Scanning electron microscope was used to determine the size, morphology, and zeta potential of nanoparticles. Flow cytometry was performed, which presented the increase percentages of CD4+ T cells (CD3e+CD4+), CD8+ T cells (CD3e+CD8+) and dendritic cells (CD11c+CD83+, CD11c+CD86+) in mice vaccinated with rHcARF1+PLGA NP. Immunoassay analysis show raised humoral (Immunoglobulin (Ig)G1, IgG2a, IgM) and cell-mediated immune response (Interleukin (IL)-4, IL-12, and IL-17, and Interferon (IFN)-γ) induced by rHcARF1+PLGA NP. Experimental groups that were treated with the antigen-loaded NP yield higher lymphocyte proliferation than the control groups. Based on these results, we could propose that the rHcARF1 encapsulated in NP could stimulate a strong immune response in mice rather than administering alone against the infection of H. contortus.

Serine/threonine-protein phosphatases (STPs), as integral constituents of parasitic excretory/secretory proteins, are assumed to be released during the host–parasite interactions. However, knowledge about these phosphatases and their immunoregulatory and immune protective efficiencies with host peripheral blood mononuclear cells (PBMCs) is scant. In this study, an open reading frame of STP from Haemonchus contortus designated as HcSTP-1 was amplified and cloned using reverse-transcription-polymerase chain reaction (RT-PCR) method. The 951-bp nucleotides sequence was encoded to a protein of 316 amino acid residues, conserved in characteristics motifs GDXHG, GDYVDRG, GNHE, HGG, RG, and H. The HcSTP-1 protein was detected at approximately 35 kDa as recombinant protein fused in an expression vector system and resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunohistochemically, HcSTP-1 was found to be localized in both male and female adult worm sections. Using immunofluorescence assay, the binding activity of rHcSTP-1 was confirmed on surface of goat PBMCs, which resulted in expression of multiple cytokines and various immunoregulatory activities in vitro. The RT-PCR results showed that mRNA level of interleukin-2, TGF-β1, IFN-γ, and IL-17 (with 10 µg/ml) was upregulated and IL-10 was decreased. However, IL-6 showed no change after PBMCs incubated with rHcSTP-1 protein. Further functional analysis showed that migratory activity of cells, intracellular nitrite production (NO), and apoptotic efficiency of PBMCs were elevated at significant level, whereas the proliferation of goat PBMCs and monocytes-associated major histocompatibility complex (MHC)-I and MHC-II expressions were decreased significantly at concentration-dependent fashion. Our results showed that the HcSTP-1 protein engaged in vital suppressive regulatory roles on host immune cells, which might represent a potential molecular target for controlling H. contortus infection in future.

Small size excretory/secretory (ES) antigens of the Haemonchus contortus parasite have intense interest among researchers for understanding the molecular basis of helminths immune regulation in term of control strategies. Immunomodulatory roles of H. contortus ES-15 kDa (HcES-15) on host immune cells during host−parasite interactions are unknown. In this study, the HcES-15 gene was cloned and expression of recombinant protein (rHcES-15) was induced by isopropyl-ß-D-thiogalactopyranoside (IPTG). Binding activity of rHcES-15 to goat peripheral blood mononuclear cells (PBMCs) was confirmed by immunofluorescence assay (IFA) and immunohistochemical analysis showed that H. contortus 15 kDa protein localized in the outer and inner structure of the adult worm, clearly indicated as the parasite’s ES antigen. The immunoregulatory role on cytokines production, cell proliferation, cell migration, nitric oxide (NO) production, apoptosis, and phagocytosis were observed by co-incubation of rHcES-15 with goat PBMCs. The results showed that cytokines IL-4, IL-10, IL-17, the production of nitric oxide (NO), PBMCs apoptosis, and monocytes phagocytosis were all elevated after cells incubated with rHcES-15 at differential protein concentrations. We also found that IFN-γ, TGF-β1, cells proliferation and migration were significantly suppressed with the interaction of rHcES-15 protein. Our findings indicated that low molecular ES antigens of H. contortus possessed discrete immunoregulatory roles, which will help to understand the mechanisms involved in immune evasion by the parasite during host−parasite interactions.

Galectins are glycan-binding proteins that are widely expressed and distributed in mammalian tissues as well as cells of innate and adaptive immune responses. CD4+ T-helper cells differentiate into effector subsets in response to cytokines. T helper 9 cells are one of the recently described subsets of effector T cells that are relatively new and less studied. In this study, galectin domain containing protein from Haemonchus contortus (Hc-GDC) was cloned, expressed in pET32a, and immunoblotting was performed. Localization of recombinant (r)Hc-GDC on outer and inner surface of H. contortus worm and binding with goat Peripheral Blood Mononuclear cells (PBMCs) were performed using immunofluorescence assay. Moreover, effects of rHc-GDC on proliferation, apoptosis, cell migration, and the nitric oxide production in goat PBMCs were evaluated. Furthermore, modulatory effects of rHc-GDC on production of Th1, Th2, and Th9 cells were evaluated by flowcytometry and on interferon gamma, interleukin (IL)-4 and IL-9 were evaluated by quantitative real-time polymerase chain reaction. The results demonstrated that rHc-GDC was successfully cloned, expressed in expression vector as well as in the gut surface of adult H. contortus worm and successful binding with PBMCs surface were observed. Immunoblotting results revealed that rHc-GDC is an important active protein of H. contortus excretory and secretory products. Moreover, the interaction of rHc-GDC with host cells increased the production of Th2, Th9 cells, IL4, IL-9, PBMC proliferation, nitric oxide, and cell migration. No effects of rHc-GDC were observed on PMBC apoptosis, production of Th1 cells, and secretions of IFN-γ and IL-10 cytokines. These findings indicate that recombinant GDC protein from H. contortus modulates the immune functions of goat PBMCs and has the potential to enhance protective immunity by inducing T helper-9-derived IL-9 in vitro.

The present study was planned to carry out a seroprevalence study of PPR antibodies in North West Frontier Province (NWFP) including Federally Administered Tribal Areas (FATA), using competitive-ELISA (c-ELISA) kit. A total of 4548 serum samples collected from 26 locations of the NWFP including. Eight hundred twenty eight samples were found positive for PPR antibodies yielding an overallseroprevalence of 18.20 percent in the small ruminant population included in the study. 337 (24.90%) serum samples from sheep and 491 (15.36%) from goats were found positive in PPR c-ELISA yielding a marginal difference in seroprevalence of PPR. The current situation of PPR in Pakistan could be regarded as of low magnitude but constant threat. Incidence may increase to the alarming level due to uncontrolled/ free movement within and from across boarders to the stocks of non-immunized susceptible population of sheep and goats in the country but production systems and economic realities constraint the effective animal movement control within and across the boarders. Based on the limited reports of outbreaks and uneven distribution of occurrence over the years of past decade, the best available options are high level of prevention and objective surveillance. Keeping in view the high risk of PPR, control strategy should be adopted and further study needs to be undertaken to ascertain the extent of PPR virus circulating in the population of goats and sheep in the country. Under existing production and marketing system, sanitation and preventive vaccination are two practicable options for control of PPR in Pakistan. Homologous PPR vaccine is now available which is a freeze dried live vaccine derived from tissue culture from attenuated PPR virus. The vaccine is intended for sheep and goats and is required to be injected to all animals above the age of 3 month. The vaccine may be repeated after one year of the primary vaccination in high risk areas. [Vet World 2009; 2(9.000): 333-336]

Haemonchus contortus is a parasitic gastrointestinal nematode, and its excretory and secretory products (HcESPs) interact extensively with the host cells. In this study, we report the interaction of proteins from HcESPs at different developmental stages to goat peripheral blood mononuclear cells (PBMCs) in vivo using liquid chromatography-tandem mass spectrometry. A total of 407 HcESPs that interacted with goat PBMCs at different time points were identified from a H. contortus protein database using SEQUEST searches. The L4 and L5 stages of H. contortus represented a higher proportion of the identified proteins compared with the early and late adult stages. Both stage-specific interacting proteins and proteins that were common to multiple stages were identified. Forty-seven interacting proteins were shared among all stages. The gene ontology (GO) distributions of the identified goat PBMC-interacting proteins were nearly identical among all developmental stages, with high representation of binding and catalytic activity. Cellular, metabolic and single-organism processes were also annotated as major biological processes, but interestingly, more proteins were annotated as localization processes at the L5 stage than at the L4 and adult stages. Based on the clustering of homologous proteins, we improved the functional annotations of un-annotated proteins identified at different developmental stages. Some unnamed H. contortus ATP-binding cassette proteins, including ADP-ribosylation factor and P-glycoprotein-9, were identified by STRING protein clustering analysis.

Background: Encephalitozoon cuniculi is a microsporidian parasite commonly found in rabbits that can infect humans, causing encephalitozoonosis. Our objective in this study was to evaluate the seroprevalence of this parasite in rabbits and humans in China. Methods: Overall, 300 serum samples each from clinically healthy rabbit and hu-man were collected from three regions of China (Sichuan Province, Chongqing Municipality and Jilin Province) from January to September 2013 and tested for anti- E. Cuniculi antibodies using an ELISA. Results: An overall seroprevalence of E. cuniculi was recorded as 56/300 (18.76%) and 29/300 (9.76%) in rabbit and human sera, respectively. The seropositivity of rabbit samples collected from Jilin province was 41%, which was significantly higher (P

The present study was conducted to check out the efficacy of various antiprotozoal drugs on bovine babesiosis, anaplasmosis and theleriosis. 38 buffaloes naturaly infected with anaplasma, babesia and theileria were selected and divided into four groups. Group A (10), B (10), C (08) and D (10) infected with anaplasmosis, babesiosis, theileriosis, and Infected Control (anaplasmosis (4), anaplasmosis + babesiosis (1 each), babesiosis (2) and theileriosis (2) respectively. Group A was further subdivided into A-I (5 animals with pure Anaplasmosis) and A-II (5 animals with mixed infection of Anaplasmosis and Babesiosis). Group-A was treated with a single dose of imidocarb dipropionate (3 mg / kg body weight). In Group A-I one animal recovered completely, 02 moderately while remaining 02 animals did not respond. The later 02 animals recovered completely after additional 02 doses of 4 mg/kg body weight. In case of group A-II complete recovery was recorded in one animal, moderate recovery in three animals while one animal showed no responsel. Group B was treated with Diminazene acceturate (3. 5 mg / kg body weight) administered as a single dose. 6 animals recovered completely, 02 moderately while 02 animals showed no response. Group C was treated with a single dose of Buparvaqune (2.5 mg / kg body weight), complete recovery was recorded in 4 animals, 2 animals showed no response, while 2 animals with severe infection died after 24 hrs. [Vet. World 2010; 3(6.000): 272-274]

Haemonchus contortus poses a global challenge as a parasite affecting small ruminants, yet the problem of absence of an effective vaccine against H. contortus infection still exists. This investigation sought to appraise the immunological reaction induced by recombinant H. contortus excretory/secretory-24 (rHcES-24) in combination with complete Freund's adjuvant (CFA) and bio-polymeric nanoparticles (NPs) within a murine model. In this study, rHcES-24 was encapsulated in poly(d, l-lactide-co-glycolide) (PLGA) and chitosan (CS) NPs, administered subcutaneously to mice. Researchers analyzed the NPs using scanning electron microscope (SEM) and assessed lymphocyte proliferation, specific antibodies, cytokines, T cell proliferation (CD3e+CD4+, CD3e+CD8a+), and phenotypic alteration in splenocytes (CD11c+CD83+, CD11c+CD86+) through flow cytometry to understand the immune response. The results demonstrated that the administration of nanovaccines (NVs) prompted immune responses towards Th1 pathway. This was indicated by notable enhancements in the production of specific antibodies, heightened cytokine levels, and a robust proliferation of lymphocytes observed in mice that received the NVs compared to control groups. Remarkably, mice vaccinated with the antigen-loaded NPs formulations exhibited considerably higher proportions of splenic dendritic cells (DCs) and T cells in comparison to those receiving the traditional adjuvant or the control groups. Incorporating HcES-24 protein into NPs effectively conferred immunity against H. contortus , paving the way for developing a targeted and commercial vaccine. [ABSTRACT FROM AUTHOR]

Background: Haemonchus contortus (H. contortus), a nematode with global prevalence, poses a major threat to the gastrointestinal health of sheep and goats. In an effort to combat this parasite, a nanovaccine was created using a recombinant ADP-ribosylation factor 1 (ARF1) antigen encapsulated within poly lactic-co-glycolic acid (PLGA). This study aimed to assess the effectiveness of this nanovaccine in providing protection against H. contortus infection. Methods: Fifteen goats were randomly divided into three groups. The experimental group received two doses of the PLGA encapsulated rHcARF1 (rHcARF1-PLGA) nanovaccine on days 0 and 14. Fourteen days after the second immunization, both the experimental and positive control groups were challenged with 8000 infective larvae (L3) of H. contortus, while the negative control group remained unvaccinated and unchallenged. At the end of the experiment on the 63rd day, all animals were humanly euthanized. Results: The results showed that the experimental group had significantly higher levels of sera IgG, IgA, and IgE antibodies, as well as increased concentrations of cytokines, such as IL-4, IL-9, IL-17, and TGF-β, compared to the negative control group after immunization. Following the L3 challenge, the experimental group exhibited a 47.5% reduction in mean eggs per gram of feces (EPG) and a 55.7% reduction in worm burden as compared to the positive control group. Conclusions: These findings indicate that the nanovaccine expressing rHcARF1 offers significant protective efficacy against H. contortus infection in goats. The results also suggest the need for more precise optimization of the antigen dose or a reassessment of the vaccination regimen. Additionally, the small sample size limits the statistical rigor and the broader applicability of the findings. [ABSTRACT FROM AUTHOR]

Bacillus thuringiensis (B. thuringiensis) is considered one of the most important entomopathogenic microorganisms. It produces potent toxins against insects. Therefore, the present study investigates the bioacaricidal properties of B. thuringiensis on the Hyalomma tick species. Firstly, we identify Hyalomma ticks based on morphological screening and molecular characterization. The cytochrome C oxidase subunit I (COX1) gene was selected for the polymerase chain reaction (PCR) analysis, which resulted in the amplification of 656 bp. The amplified products were sequenced, and the isolated (COX1) gene of ticks was submitted to the gene bank of NCBI (Accession No. OR077934.1). The nucleotide sequences were retrieved from the NCBI data bank by BLASTn analysis, which confirmed that all obtained sequences belong to genus Hyalomma, and multiple alignments confirmed that the sequence of Hyalomma anatolicum Tandojam‐isolate (HA‐TJ) 100% aligned with Hyalomma analoticum KP792577.1, Hyalomma detritum KP792595.1, Hyalomma excavatum KX911989.1, and H. excavatum OQ449693.1. The generated phylogenetic tree confirmed that sequences of HA‐TJ COX1 clustered with a single clad of H. analoticum, H. excavatum, and H. detritum. The acaricidal effect of B. thuringiensis toxins B. thuringiensis spore crystal mix (BtSCM) and B. thuringiensis crystal proteins (Btcps) was evaluated against larvae and adult life stages of Hyalomma ticks in vitro. We applied Btcps and BtSCM separately with different concentrations and calculated the mortality percentage. Adult mortality was estimated at the 8th, 10th, 12th, and 15th days posttreatment and larval mortality after 24 h. During treatment of the adult life stage, at first, ticks were immersed in different concentrations of Btcps and BtSCM for 5 min after the treatments, and the samples were transferred to sterile containers and placed in an incubator with 80% humidity at 23°C. Furthermore, Btcps produced the highest mortality on Day 15, 89 ± 1.00% at a concentration of 3000 μg/mL, followed by the 12th, 10th, and 8th days produced 83 ± 1.91%, 70 ± 1.15%, and 61 ± 1.00%, respectively. BtSCM produced mortality of 69 ± 1.91% on Day 15 at a concentration of 3000 μg/mL, followed by the 12th, 10th, and 8th days at 57 ± 2.51%, 37 ± 1.91%, and 34 ± 2.00%. The present study revealed that B. thuringiensis toxins produced a significant (p < 0.05) increase in mortality rate in adults of Hyalomma ticks. Additionally, Btcps and BtSCM were used to treat the larval stage. The treatments were applied to calculate the mortality percentage via the Laravel packet test. At a 1500 μg/mL concentration, Btcps resulted in the highest mortality of 98 ± 1.15%; this was followed by 1250 μg/mL, 1000 μg/mL, and 750 μg/mL, which produced mortalities of 76 ± 1.63%, 60 ± 1.63%, and 56 ± 1.63%, respectively. In addition, BtSCM produced a mortality rate of 79 ± 2.51% at a concentration of 1500 μg/mL. Furthermore, 75 ± 2.51%, 65 ± 1.91%, and 58 ± 1.15% mortality were observed at concentrations of 1250 μg/mL, 1000 μg/mL, and 750 μg/mL, respectively. The results showed a significant (p < 0.05) increase in larval mortality compared to the control group. We conclude that B. thuringiensis toxins are applicable as a bioacaricide. [ABSTRACT FROM AUTHOR]