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Introduction: Toxoplasmosis is caused by the opportunistic, cosmopolitan protozoan Toxoplasma gondii is one of the most common parasitoses in the world. This parasite can pose a threat to people with immunodeficiency but also to the fetus, since the invasion can lead to miscarriages. Moreover, this parasite can contribute to economic losses in livestock farming. These problems lead to the implementation of new, safe solutions for the development of effective toxoplasmosis immunoprophylaxis. Methods: In this work, newly produced recombinant trivalent chimeric proteins of T. gondii , based on SAG1-SAG2 recombinant chimeric antigen that differ in one terminal antigenic component, were tested in terms of their ability to induce an effective post-vaccination response. Antigens were tested in vitro to assess their ability to elicit APC cells response and further mice of the C3H/HeOuJ strain were immunized using those antigens, to evaluate their immunogenicity and immunoprotective effect in vivo. Two weeks after the last dose mice were either sacrificed to assess selected parameters of the immune response or infected with T. gondii DX strain to determine the degree of protection one month later. Results: The results of serological tests revealed a high level of serum IgG antibodies specific for the native T. gondii TLA antigens. TLA-stimulated splenocytes produced cytokines that are important in inhibiting protozoal invasion. Additionally, CD3+ CD4+ and CD3+ CD8+ T cell subpopulations of splenocytes were analysed by flow cytometry. One month after experimental infection mice were sacrificed, and their brains were isolated to count T. gondii tissue cyst. Immunization of mice with recombinant trivalent chimeric proteins of T. gondii resulted in reduction of tissue cyst burden rates reaching even 74%. Discussion: The obtained results demonstrate strong immunogenicity of the studied proteins and will allow to select candidates for further research aimed at increasing the immunoprotective properties of experimental vaccines against toxoplasmosis based on T. gondii chimeric antigens. [ABSTRACT FROM AUTHOR]

Vector‐borne diseases represent various sicknesses that are increasingly significant in human and veterinary health. Among the zoonotic agents transmitted by ticks, infections caused by the intracellular pathogens from the Anaplasmataceae family and piroplasmids (Babesia/Theileria spp.) are particularly notable due to the substantial economic losses they cause in the livestock sector. A study was carried out to assess the prevalence of these pathogens in ticks from ruminants in the province of Valencia (Eastern Iberian Peninsula). Between 2019 and 2022, 1674 ticks were collected from 163 domestic and wild ruminants. The tick species were identified using molecular and morphological features and included Rhipicephalus spp. (Rhipicephalus bursa and Rhipicephalus sanguineus) (88.7%), Haemaphysalis sulcata (5.9%), Ixodes ricinus (3.6%) and Dermacentor marginatum (1.2%). The Spanish ibex (Capra pyrenaica) was found to be at greater risk of infestation by Haemaphysalis compared to other ruminants. Polimerase Chain Reaction (PCR) analysis was performed on 108 individual ticks from different hosts, with 16.7% testing positive for members of the Anaplasmataceae family and 39.8% for Theileria spp. Sequencing of 10 positive samples from each pathogen group revealed the presence of Anaplasma ovis, Anaplasma platys, Ehrlichia spp., Theileria ovis and Theileria capreoli. No Anaplasma phagocytophilum or Babesia spp. were detected. Further research is essential for improved management of ticks and the diseases they transmit in the study area. [ABSTRACT FROM AUTHOR]

This study designs a theoretical model and simulation model that can explain the welfare impacts of disease transmission that occurs in livestock trade. A household production model and a SIR model are used to find theoretical profitable conditions for infectious livestock trading and prices and quantities for transactions. Under the theoretical conditions an agent-based model is used to simulate livestock transactions to compare social impacts based on the number of livestock, household wealth and income, and wealth inequality. Asymmetric information is used to assign tendencies of livestock trading agents. Buyers are assumed to be uninformed about the health status of livestock owned or used by sellers, while sellers are either uninformed for their herd's health status, and if informed, the sellers' behavior of selecting infectious livestock for transactions is divided into selfish selection and altruistic selection. The simulation results reveal that livestock losses are higher when trading occurs, but overall economic welfare tends to increase with trade. Interestingly, when sellers selfishly sell sick animals, average household wealth and income peak, albeit with greater wealth inequality. [ABSTRACT FROM AUTHOR]

Large carnivore species frequently predate and consume wild or domestic prey, which is referred to as food-related predation. Large carnivores can also hunt and kill prey exceeding their immediate needs (i.e., they do not consume prey), which is referred to as surplus predation. We used 173 records of livestock predations by gray wolves (Canis lupus; n = 133) and Persian leopards (Panthera pardus tulliana; n = 40) reported by governmental organizations of Iran during 2009–2019 to investigate food-related and surplus predation incidents of livestock. We found that for wolves, the number of reported surplus predation incidents was greater than that of food-related predation incidents during all 4 seasons (spring through winter), whereas for leopards, the number of food-related and surplus predation incidents were similar in all seasons. The number of livestock killed per surplus predation incident was greater for wolves than for leopards and that surplus predations by both species occurred more frequently within corrals than on free-range pastures. As corrals in most villages across Iran are poorly constructed and largely accessible to predators, we recommend that livestock owners enhance corral construction, use well-trained dogs during day and particularly at night, employ people to watch livestock at night, and use fire (e.g., torches) during night to scare carnivores. These strategies can mitigate predation incidents and corresponding economic losses, resulting in fewer losses of livestock, wolves, and leopards, as these two carnivore species are mainly killed by humans due to livestock predations across Iran. [ABSTRACT FROM AUTHOR]

Brucellosis is a serious zoonosis caused by Brucella spp. infection, which not only seriously jeopardizes the health of humans and mammals, but also causes huge economic losses to the livestock industry. Brucella is a Gram-negative intracellular bacterium that relies primarily on its virulence factors and a variety of evolved survival strategies to replicate and proliferate within cells. Currently, the mechanisms of autophagy, inflammation, and apoptosis in Brucella -infected hosts are not fully understood and require further research and discussion. This review focuses on the relationship between Brucella and autophagy, inflammation, and apoptosis to provide the scientific basis for revealing the pathogenesis of Brucella. [ABSTRACT FROM AUTHOR]

Porcine epidemic diarrhea (PED) has caused serious economic losses to the swine livestock industry. Due to the rapid variation in the PEDV) genome, especially the spike (S) protein, the cross-protection ability of antibodies between different vaccine strains is weakened. Hence, the rapid development of safe, broad-spectrum and highly effective attenuated PEDV vaccine still needs further research. Here, we found that the replacement of the S2 subunit had little effect on S protein immunogenicity. Moreover, the chimeric virus (YN-S2DR13), the S protein of the YN strain was replaced by the DR13 S2 subunit, which lost its trypsin tropism and increased its propagation ability (approximately 1 titer) in Vero cells. Then, the pathogenesis of YN-S2DR13 was evaluated in neonatal piglets. Importantly, quantitative real-time PCR, histopathology, and immunohistochemistry confirmed that the virulence of YN-S2DR13 was significantly reduced compared with that of YN. Immunization with YN-S2DR13 induced neutralizing antibodies against both YN and DR13 in weaned piglets. In vitro passaging data also showed that YN-S2DR13 had good genetic stability. Collectively, these results suggest that YN-S2DR13 has significant advantages as a novel vaccine candidate, including a capacity for viral propagation to high titers with no trypsin requirement and the potential to provide protection against both PEDV G1 and G2 strains infections. Our results also suggests that S2 subunit replacement using reverse genetics can be a rapid strategy for the rational design of live attenuated vaccines for PEDV. IMPORTANCE Emerging highly virulent porcine epidemic diarrhea virus (PEDV) G2 strains has caused substantial economic losses worldwide. Vaccination with a live attenuated vaccine is a promising method to prevent and control PED because it can induce a strong immune response (including T- and B-cell immunity). Previous studies have demonstrated that the S2 subunit of the PEDV spike (S) protein is the determinant of PEDV trypsin independence. Here, we evaluated the pathogenicity, tissue tropism, and immunogenicity of the chimeric virus (YN-S2DR13) via animal experiments. We demonstrated that YN-S2DR13 strain, as a trypsin independent strain, increased intracellular proliferation capacity, significantly reduced virulence, and induced broad-spectrum neutralization protection against PEDV G1 and G2 strains. In vitro passaging data also validated the stability of YN-S2DR13. Our results showed that generating a chimeric PEDV strain that is trypsin-independent by replacing the S2 subunit is a promising approach for designing a live attenuated vaccine for PEDV in the future. [ABSTRACT FROM AUTHOR]

In the Alpine region, the use of herd protection measures is assumed to not be reasonable in many places due to geographical, technical and economic limitations. In line with EU regulations, securing compensation for livestock damage or losses necessitates evidence of the feasibility of these measures. The aim of this study was to establish a scientifically validated classification framework for assessing the viability of employing herd protection measures (livestock guarding fences and guarding dog utilization) in alpine pastures. To achieve this, five representative Alps in the Italian alpine province of South Tyrol were considered using a devised classification framework and geospatial data. The resultant outcomes were validated through on-site inspections. Six categories were identified for the use of livestock guarding fences (proximity to roads, herding situation, topography, tourism, pasture geometry and proportion of not fencable area) and, in addition to these, two categories (herd homogeneity and number of herded livestock) for the use of livestock guarding dog utilization. The validation showed that the basic assumptions are plausible, but that the geodata partly show inaccuracies in the demarcation of hazard zones, hiking trails and pasture geometry. Despite these limitations, the developed classification framework was suitable for classifying the practicability of herd protection measures on alpine pastures. Therefore, it provides the necessary factual basis and logical reasoning for legally secure compensation payments. Additionally, our classification framework sets the stage for considering strategic predator management approaches. [ABSTRACT FROM AUTHOR]

Background/Objectives: Bovine babesiosis is a vector-borne disease transmitted by ticks that causes important losses in livestock worldwide. Recent research performed on the drugs currently used to control bovine babesiosis reported several issues including drug resistance, toxicity impact, and residues in edible tissue, suggesting the need for developing novel effective therapies. The endochin-like quinolones ELQ-316 and buparvaquone (BPQ) act as cytochrome bc1 inhibitors and have been proven to be safe and efficacious against related apicomplexans, such as Plasmodium spp. and Babesia microti, without showing toxicity in mammals. The objectives of this study are investigating whether ELQ-316, BPQ, and their combination treatment could be effective against Babesia bovis in an in vitro culture model and comparing with imidocarb (ID), the routinely used drug. Methods: In vitro cultured parasites starting at 2% percentage of parasitemia (PPE) were treated with BPQ, ELQ-316, ID, and the combinations of BPQ + ELQ-316 and ID + ELQ-316 at drug concentrations that ranged from 25 to 1200 nM, during four consecutive days. The IC50% and IC99% were reported. Parasitemia levels were evaluated daily using microscopic examination. Data were compared using the non-parametrical Mann–Whitney and Kruskall–Wallis test. Results: All drugs tested, whether used alone or in combination, significantly decreased the survival (p < 0.05) of B. bovis in in vitro cultures. The combination of BPQ + ELQ-316 had the lowest calculated inhibitory concentration 50% (IC50%) values, 31.21 nM (IC95%: 15.06–68.48); followed by BPQ, 77.06 nM (IC95%: 70.16–86.01); ID + ELQ316, 197 nM (IC95%:129.0–311.2); ID, 635.1 nM (IC95%: 280.9–2119); and ELQ316, 654.9 nM (IC95%: 362.3–1411). Conclusions: The results reinforce the higher efficacy of BPQ at affecting B. bovis survival and the potential synergistic effects of its combination with ELQ-316, providing a promising treatment option against B. bovis. [ABSTRACT FROM AUTHOR]

Importance: Gastrointestinal nematodiasis poses significant economic losses in the livestock industry due to mortality, morbidity, and decreased production. Objective: This study examined the prevalence of gastrointestinal nematodiasis in dairy cattle in Central Inner Mongolia, Northern China, the associated risk factors, and the effects on the pepsinogen and gastrin levels. Methods: Fecal samples (n = 590) were collected from four regions and analyzed using the standard floatation and sedimentation techniques. The serum gastrin and pepsinogen levels were also assessed as potential biomarkers for gastrointestinal nematodiasis. Results: Among 590 animals, 259 (43.9%; 95% confidence interval, 36.7–49.8) tested positive for at least one type of gastrointestinal nematode parasite. Among the positive animals, 38.6% were poly-parasitic, while 61.4% were mono-parasitic (p < 0.05). Cooperia was the predominant nematode among mono-infections, accounting for 10.8%, followed in order by Oesophagostomum (10.0%), Trichuris (9.6%), Trichostrongylus (8.5%), Dictyocaulus (6.9%), Ascarid (6.9%), Haemonchus (4.6%), and Strongyloides (3.9%). Among the poly-parasitic infected cattle, 22.7%, 9.3%, 4.2%, and 2.3% had two, three, four, and five parasite combinations, respectively. The risk factors, such as age, breed, area, physical condition, and presence of diarrhea, were significantly (p < 0.05) associated with the prevalence of gastrointestinal nematodiasis. Poly-parasitic infected cattle showed 49.1% higher (p = 0.013) serum pepsinogen levels than the control group, while the mono-parasitic ones exhibited a 28.33% increase (p = 0.030). Similarly, the serum gastrin levels increased by 99.2% (p = 0.004) and 71.3% (p = 0.015) in the poly-parasitic and mono-parasitic animals, respectively. Conclusions and Relevance: These findings demonstrate the high prevalence of gastrointestinal nematodiasis in Northern China, with an alarming rate of poly-parasitic infections. In addition, polyparasitic infections affect the enzymes pepsinogen and gastrin levels. [ABSTRACT FROM AUTHOR]

Mitigation practices for nitrogen leaching losses from livestock agriculture are needed to protect freshwater quality and increase the efficiency of agricultural production. Within New Zealand, the most common pasture type is a twospecies mix of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens). Ecological theory suggests that increasing species and functional diversity improves ecosystem function, including nitrogen (N) retention. Use of more diverse pasture types, including a mix of pasture grasses, legumes and other forbs, particularly plantain (Plantago lanceolata), with functional traits, including winter activity, deep-rooting, N fixation, and biological inhibition of nitrification in the soil, is a potential mitigation practice that requires further verification with long-term field measurements. Here we utilize a network of large lysimeters to make field-based measurements of N leaching from 5-8 species diverse pasture, including plantain, under a range of soil, climate and management conditions, for comparison with losses from traditional ryegrass-clover pasture. Over 3 years of measurements, leaching from fully established diverse pasture was 2-80 kg N ha-1 y-1. No differences were observed in dry matter production or N leaching of diverse pasture compared to ryegrass-clover lysimeters. Large losses, up to 120 kg N ha-1, were observed during periods when pasture was not fully established, including cultivation and sowing of new pasture, depending on season. Timing of management activities could be optimized to minimize these losses. These data provide critical assessment of diverse pasture as a mitigation approach for reducing N losses. Further work on diverse pastures should include higher diversity mixes as well as consideration of animal mediated effects of diverse pasture diets on N inputs. [ABSTRACT FROM AUTHOR]

Background: Toxoplasmosis not only leads to abortion in humans but also in herbivores, which causes significant financial and quality-adjusted life-year losses. The present study aimed to determine the prevalence of toxoplasmosis in aborted fetuses via serological and molecular assays. Moreover, the genotypes of the obtained isolates were detected. Methods: Serological and molecular methods were used to study aborted fetuses from Bojnourd City, North Khorasan Province, Iran, which included 52 ovines and 16 bovines. Nested PCR of the B1 gene was used to detect parasite DNA in brain tissues. The PCR-RFLP method for the GRA6 gene was used to determine the genotype of T. gondii. Results: Out of 68 aborted fetuses, 16.1% showed the presence of anti-T. gondii IgG. Among these, 11.7% were identified in bovine fetuses and 4.4% in ovine fetuses. Additionally, two (2.94%) samples of ovine tested positive for anti-T. gondii IgM. Our PCR analysis detected parasite DNA in two cases (2.94%) among 11 IgG-positive samples. All obtained isolates belong to type I of T. gondii. Conclusion: Infection with Type I of T. gondii during the neonatal period may partly be responsible for abortion and economic losses in livestock farming in our studied region. To understand the molecular epidemiology and genotypes of T. gondii associated with abortion, further evaluation of aborted samples from different geographical locations is necessary. [ABSTRACT FROM AUTHOR]

Simple Summary: House flies are important carriers of diseases and can cause economic losses in livestock by reducing feeding activity. Overuse of chemical insecticides has led to resistance in many house fly populations worldwide. This study aimed to assess how two house fly populations in Antalya, Türkiye respond to the insecticides cyfluthrin and etofenprox. By exposing adult flies to these chemicals, the researcher found that the Kemer population showed very low resistance to cyfluthrin, while the Serik population exhibited moderate resistance. For etofenprox, Kemer flies showed no resistance, and Serik flies showed very low resistance. The findings suggest that integrated pest control methods and regular resistance monitoring are necessary to manage and prevent further resistance in house fly populations. The house fly, Musca domestica L. (Diptera: Muscidae), is a significant vector for many pathogens and parasites. Presence of this vector causes economic losses due to decreased feeding activity in livestock, resulting in reduced yields of products. The repeated and unconscious application of chemical insecticides has resulted in the development of resistance in the majority of house fly populations worldwide. The aim of this research was to determine the susceptibility levels of two field M. domestica populations to cyfluthrin and etofenprox in Antalya, Türkiye. The LD50 values, resistance ratios, and resistance status were determined by exposing adult house flies to the test chemicals for one hour. The Kemer population exhibited very low resistance to cyfluthrin, with a 5-fold increase, while the Serik population, in contrast, showed moderate resistance with a 29.67-fold increase. The resistance ratios of field populations to etofenprox were 2.33-fold (no resistance) for Kemer and 6.44-fold (very low resistance) for Serik, respectively. This is the first study to determine the resistance levels of house flies against cyfluthrin and etofenprox insecticides in Türkiye. To prevent or reduce the development of resistance to insecticides in house flies, integrated control methods and resistance management programs should be employed. Furthermore, it is advised that regular monitoring tests be conducted to observe the level of resistance. [ABSTRACT FROM AUTHOR]

Lumpy skin disease (LSD) poses a significant threat to animal welfare and leads to economic losses in affected countries. LSD outbreaks may also impact trade and regional relationships. South and Southeast Asia, with its dense livestock population and intricate trade networks, are susceptible to LSD outbreaks. Indonesia confirmed its first LSD cases in March 2022, leading to substantial livestock losses by August 2023. Australia, an important player in the global beef industry, faced trade disruptions due to LSD concerns raised by Indonesia and Malaysia, claims that were refuted by Australian authorities. The dispute highlights the need for good surveillance. EPIWATCH®, employing artificial intelligence, provides real‐time outbreak signals, and spatial analysis can identify LSD hotspots, leading to timely interventions. This study uses data collected by the EPIWATCH® open‐source disease surveillance system at the University of New South Wales in 2022 and 2023 and compares it for timeliness and completeness with data available on the World Animal Health Information System (WAHIS). We found more timely reports of LSD outbreaks in EPIWATCH® compared to WAHIS. In conclusion, open‐source surveillance tools like EPIWATCH® can provide timely alerts of disease emergence, such as LSD outbreaks in South and Southeast Asia, which can supplement formal reporting systems. [ABSTRACT FROM AUTHOR]

Ticks are responsible for transmitting of pathogenic microorganisms during their feeding process on the hosts. They also cause significant losses in livestock production and, in many cases, the death of infected animals. In recent decades, many efforts have been carried out to combat ticks by using natural compounds. The present study aimed to evaluate the acaricidal effect of the hydroalcoholic extract of honey bee propolis against Haemaphysalis spp. in vitro. The acaricidal activities of the propolis were considered at concentrations of 25, 50, and 100 mg/ml and negative and positive controls (distilled water and Cypermethrin) following 10, 30, and 60 minutes of exposure. In this experiment the spraying and contact methods were used, and all tests were repeated twice. The chemical composition of propolis was identification by Gas Chromatography-Mass Spectrometry (GC–MS). Data were analyzed using GraphPad Prism software version 5.0. According to the results, propolis had an acaricidal effect; however, this effect was more potent in the spraying. The propolis showed a 100% mortality rate at 100 mg/ml concentrations after 60 min exposure. GC-MS investigation showed that Heptanone (48.65%) was the main ingredient of propolis. The results indicated that the hydroalcoholic propolis extract carry potent acaricidal ingredients and might afford new natural acaricidal compounds for the control of Haemaphysalis spp. [ABSTRACT FROM AUTHOR]

The Rhipicephalus microplus tick causes enormous economic losses in livestock farming around the world. Despite several promising studies carried out with plant extracts such as Achyrocline satureioides against this ectoparasite, a major obstacle is related to pharmaceutical presentation forms. There is no study showing xantan gum-based hydrogel and polycaprolactone nanoparticles containing A. satureioides extract against R. microplus larvae. The objective of this study was to incorporate A. satureioides extract to develop a nanoformulation (AScn) and a hydrogel (ASlh) and evaluate them against R. microplus larvae with the purpose of increasing the contact time of the extract with the larvae and improve the effectiveness. The ethanolic extracts were incorporated in polycaprolactone nanoparticles and characterized via analysis of the mean hydrodinamic diameter and polidispersity index. The xanthan gum-based hydrogel formulation was prepared with crude extract of A. satureioides 40 mg/mL, 0.25% xanthan gum, and 8% poloxamer, to determine the bioadhesiveness of the formulation in bovine leather and the flow rate of the formulation in the animal. The results in larvae demonstrated that when evaluated in the form of a hydrogel (ASlh), mortality was higher, with 91.48% mortality at a concentration of 20 mg/mL presenting itself as an interesting alternative for controlling this ectoparasite. [ABSTRACT FROM AUTHOR]

Lameness is one of the most common health issues in cattle, which is a significant cause of health disorders and economic losses in livestock. Due to the numerous and varied causes, its treatment and prevention are complex and challenging. Lameness is associated with intense pain, leading to discomfort, reduced milk yield, weight loss, and reproductive issues, which can ultimately result in chronic lameness and substantial financial losses. A number of simple, visual scoring scales have been developed for their detection and evaluation, which enable them to be detected quickly on the one hand, and appropriate interventions to be taken and their effectiveness evaluated on the other. The consistent application of these scoring systems is becoming a critical component of herd management strategies, aimed not only at reducing the prevalence of lameness but also at minimizing the associated economic impacts. This dual focus-improving animal welfare and reducing financial losses-highlights their importance in maintaining both herd health and cost-effectiveness. [ABSTRACT FROM AUTHOR]

Simple Summary: The infection of animals and humans with Sarcocystis spp. causes public health problems and significant economic losses to the livestock industry. To date, more than 200 species in the genus Sarcocystis have been reported. Sarcocystis spp. can cause sarcocystosis, which leads to emaciation, abortion, and even death in sheep. However, to date, Sarcocystis spp. infection has not been reported in sheep (Ovis aries) in Shanxi Province. In the present study, 582 mutton samples collected from sheep in ten representative counties in this province were investigated for Sarcocystis spp. by PCR amplification of the mitochondrial cytochrome c oxidase subunit I (cox1) gene. The results showed that the prevalence of Sarcocystis spp. in sheep in Shanxi Province was 33.85% (197/582). Of them, 196 cox1 sequences showed a nucleotide similarity of 98.56–99.81% with those of S. tenella, and the remaining one cox1 sequence showed a nucleotide similarity of 99.71% with that of S. arieticanis. The present study reported the occurrence and prevalence of Sarcocystis spp. for the first time in sheep in Shanxi Province, China, which has important implications for the control and prevention of Sarcocystis infection in this province and elsewhere. Sarcocystis species are intracellular coccidian protozoans that can infect a range of animals and humans and cause public health problems and economically significant losses. Sarcocystosis in sheep (Ovis aries) can cause abortion, neurological symptoms, and even death and results in significant economic losses to the livestock industry. To date, however, it is yet unknown whether sheep in Shanxi Province, north China, are infected with Sarcocystis spp. The purpose of this study was to investigate the prevalence of Sarcocystis spp. in sheep in Shanxi Province. Thus, 582 muscle samples of sheep were purchased from farmers' markets from ten representative counties in Shanxi Province, north China, and examined for the presence and prevalence of Sarcocystis spp. by PCR amplification of the mitochondrial cytochrome c oxidase subunit I (cox1) gene. Of the examined 582 mutton samples, 197 samples (33.85%) were Sarcocystis-positive and were sequenced. Of the obtained 197 cox1 sequences, 196 sequences showed nucleotide similarity of 98.56–99.81% with those of S. tenella, and the remaining one cox1 sequence showed nucleotide similarity of 99.71% with that of S. arieticanis. Two representative cox1 sequences of S. tenella (accession nos. PQ189447 and PQ189448) have 99.52% and 99.61% identity with S. tenalla (KC209725) and S. tenalla (MK419984), respectively. The sequence of S. arieticanis (accession no. PQ165949) obtained in this study has 99.71% identity with S. arieticanis (MK419975). This present study documents the occurrence and prevalence of Sarcocystis spp. in sheep in Shanxi Province for the first time, which enriches the data on the distribution of Sarcocystis spp. in sheep in China and has implications for the control of sheep sarcocystosis. [ABSTRACT FROM AUTHOR]

Background: Colibacillosis caused by Escherichia coli causes significant economic losses in the livestock sector worldwide and is one of the calves' leading causes of diarrhea. Aim: This study aimed to identify the most frequent E. coli molecularly pathotypes in calves with diarrhea in six provinces of the Cajamarca region in the northern highlands of Peru. Methods: Twenty-eight herds of dairy cattle under a semi-intensive rearing system were evaluated; 95 samples were isolated from calves with diarrhea up to the first month of life, 62 males and 33 females, during the rainy season. Results: The presence of virulence genes of E. coli strains was more prevalent in males; the astA (89.47%), st (83.15%), and f5 (57.89%) genes were more expressed, and the lt (17.89%) and stx2 (1.05%) genes were less expressed. The eae gene (21.05%) was more present in females. Conclusion: When E. coli strains express virulence genes astA, st, and f5 and their atypical double, triple, and quadruple combination between different observed pathotypes, they give rise to the formation of several pathotypes by the horizontal transfer of virulence genes, which can cause colibacillosis processes in more virulent calves, which is one of the most important causes of diarrhea in calves in the region of Cajamarca, compromising the sanitary viability in the herds. [ABSTRACT FROM AUTHOR]

Sheep caseous lymphadenitis (CLA) causes significant economic losses in the livestock sector by causing a loss in the quantity and quality of animal products and a loss in the breeding value of animals. Although the primary agent in CLA's etiology is Corynebacterium pseudotuberculosis, some other opportunistic microorganisms also play a role. Therefore, the control and treatment of CLA necessitates the identification of the relevant etiological agents. This study aimed to conduct an in vitro culture and molecular characterization (PCR analysis and 16S rRNA sequencing) of the bacteria involved in sheep CLA cases reported in the Çankırı province of Türkiye and determine the antibiotic susceptibility of the case isolates. In total, 82 (16.4%) of 500 sheep in five farms were diagnosed with CLA. Following the culture of the superficial abscesses samples, C. pseudotuberculosis was identified in 30 (36.59%) as a result of PCR, Pseudomonas spp. in 8 (9.76%), and Enterobacter cancerogenus in 1 (1.22%), as a result of 16S rRNA sequencing. These data revealed extensive heterogeneity among the Pseudomonas isolates, with hints of derivation from a common ancestry for some and phylogenetic similarity to isolates from Germany, Malaysia, and India. In contrast to the high susceptibility to cefoperazone and lincomycin, the high resistances of C. pseudotuberculosis and Pseudomonas spp. isolates to cephalothin, ceftiofur, cloxacillin, amoxicillin, and bacitracin were remarkable. Based on these findings, it was concluded that for an effective treatment and control of ovine CLA cases, there is a need to consider the possible involvement of opportunistic bacteria other than the primary causative agent, C. pseudotuberculosis. It also contributed to increasing the country-specific sequence data and establishing new taxa from a universal perspective. [ABSTRACT FROM AUTHOR]

Tropical theileriosis is a lymphoproliferative disease caused by Theileria annulata and is transmitted by Ixodid ticks of the genus Hyalomma. It causes significant losses in livestock, especially in exotic cattle. The existing methods for controlling it, chemotherapeutic agents and a vaccine based on an attenuated schizont stage parasite, have several limitations. A promising solution to control this disease is the use of molecular vaccines based on potential immunogenic proteins of T. annulata. For this purpose, we selected five antigenic sequences of T. annulata, i.e. SPAG-1, Tams, TaSP, spm2, and Ta9. These were subjected to epitope prediction for cytotoxic T lymphocytes, B-cells, and helper T lymphocytes. CTL and B-cell epitopes with a higher score whereas those of HTL with a lower score, were selected for the construct. A single protein was constructed using specific linkers and evaluated for high antigenicity and low allergenicity. The construct was acidic, hydrophobic, and thermostable in nature. Secondary and tertiary structures of this construct were drawn using the PSIPRED and RaptorX servers, respectively. A Ramachandran plot showed a high percentage of residues in this construct in favorable, allowed, and general regions. Molecular docking studies suggested that the complex was stable and our construct could potentially be a good candidate for immunization trials. Furthermore, we successfully cloned it into the pET-28a plasmid and transformed it into the BL21 strain. A restriction analysis was performed to confirm the transformation of our plasmid. After expression and purification, recombinant protein of 49 kDa was confirmed by western blotting. An ELISA detected increased specific antibody levels in the sera of the immunized animals compared with the control group, and flow cytometric analysis showed a stronger cell-mediated immune response. We believe our multi-epitope recombinant protein has the potential for the large-scale application for disease prevention globally in the bovine population. This study will act as a model for similar parasitic challenges. [ABSTRACT FROM AUTHOR]

Brucellosis is a zoonosis caused by bacteria of the genus Brucella, which results in economic losses relating to livestock and threatens public health. A cross-sectional study was conducted to determine the molecular prevalence of Brucella species in smallholder dairy cattle in six regions of Tanzania from July 2019 to October 2020. Dairy cattle (n = 2048) were sampled from 1371 farms. DNA extracted from blood and vaginal swabs was tested for Brucella using qPCR targeting the IS711 gene and positives were tested for the alkB marker for B. abortus and BMEI1172 marker for B. melitensis. The molecular prevalence was 3.5% (95% CI: 2.8–4.4) with the highest prevalence 8.1% (95% CI: 4.6–13.0) in Njombe region. B. melitensis was the predominant species detected (66.2%). Further studies are recommended to understand the source of B. melitensis and its implications for veterinary public health. Livestock keepers should be informed of the risks and biosecurity practices to reduce the introduction and control of Brucella. Cattle and small ruminant vaccination programs could be implemented to control brucellosis in high-risk populations in the country. [ABSTRACT FROM AUTHOR]

Benefit-sharing mechanisms have been instrumental in securing the support of local communities living on the edge of protected areas, and in resolving human-wildlife conflicts. Understanding how current conservation policies influence benefit-sharing and the management of human-wildlife conflicts is vital for designing strong mechanisms to achieve both conservation and ethical community outcomes. Here we examined how benefit-sharing and human-wildlife conflicts are acknowledged and addressed within the current conservation policies (wildlife policies, legislations, regulations and strategic plans) using the Greater Serengeti Ecosystem (GSE) as a case study. The fourteen policies focusing primarily on wildlife conservation in Tanzania are analysed. We also drew on the findings of three studies conducted during 2021 and 2022 documenting community interactions with wildlife and adjacent protected areas. These studies covered the types of benefit provided by various conservation institutions to local communities, the effectiveness of benefit-sharing mechanisms, and the conservation-related losses incurred by communities. The study uncovered a wide range of benefits provided to local communities in the form of social amenities, livelihood support and employment. However, the losses and costs experienced are far greater than these benefits. Integrating these findings with policy analysis, we identified that the main limitations reducing the impacts of benefits in motivating positive conservation behaviours to be: losses of livestock and crops were more than four times the value of benefits/compensation offered to communities; a lack of details and clarity of benefit-sharing mechanisms; benefit/conservation rates that have not been indexed against inflation and; shortcomings in implementation. To achieve conservation outcomes, we recommend revising the conservation policies and legislation across the country in collaboration with local communities who experience the impacts of conservation in their day-to-day life. We also propose a coordinated approach for all conservation institutions managing wildlife to address widespread human-wildlife conflicts across the country effectively and to achieve better conservation outcomes rather than leaving that task to a single organization (i.e. Tanzania Wildlife Management Authority). [ABSTRACT FROM AUTHOR]

In many developing countries, poor households face multiple shocks that disrupt their consumption patterns and lead to an increased welfare loss. Consequently, households adopt coping mechanisms that negatively impact their overall well-being. While social safety nets have been implemented to protect vulnerable households, it is essential to assess their overall effectiveness, both in terms of their impact and their potential to replace the harmful coping strategies adopted by households. Therefore, the primary objective of this study is to examine the impact of cash transfers provided by the Benazir Income Support Program (BISP) on the consumption patterns of households in Pakistan, with a particular emphasis on how these transfers provide consumption insurance to ultra-poor families encountered with idiosyncratic and covariate shocks. The study analyzes both food and non-food expenditure to gain insights into the effectiveness of the program in providing economic security to vulnerable households. Using three rounds of the BISP survey (2011–2013–2016), the empirical analysis is done by employing regression discontinuity design (RDD), difference-in-difference (DID) technique, and ordinary least squares method. Using the DID approach, we find that from 2011 to 2016 the overall food and non-food consumption of BISP beneficiaries have increased considerably and these results are highly significant as well. However, the estimates for RDD are not significant for the years 2013 and 2016. We also observed households' consumption behavior in the presence of shocks and found that in the wake of idiosyncratic shocks, poor households can protect their consumption through informal transfer mechanisms by securing loans from family, friends, and landlords. However, this mechanism collapses when households encounter covariate shocks. Furthermore, BISP cash transfers are primarily effective in mitigating the impact of employment loss, loss of livestock, and the rise in food prices and remain inadequate in providing insurance against major other shocks encountered by the targeted poor households. The results indicate that BISP has the potential to substitute informal coping strategies used by the poor, it requires substantial transfers to offer comprehensive consumption insurance to the poorest of the poor in Pakistan. [ABSTRACT FROM AUTHOR]

Brucellosis is a chronic and debilitating disease in humans, causing great economic losses in the livestock industry. Making an effective vaccine is one of the most important concerns for this disease. The new mRNA vaccine technology due to its accuracy and high efficiency has given promising results in various diseases. The objective of this research was to create a novel mRNA vaccine with multiple epitopes targeting Brucella melitensis. Seventeen antigenic proteins and their appropriate epitopes were selected with immunoinformatic tools and surveyed in terms of toxicity, allergenicity, and homology. Then, their presentation and identification by MHC cells and other immune cells were checked with valid tools such as molecular docking, and a multi-epitope protein was modeled, and after optimization, mRNA was analyzed in terms of structure and stability. Ultimately, the immune system's reaction to this novel vaccine was evaluated and the results disclosed that the designed mRNA construct can be an effective and promising vaccine that requires laboratory and clinical trials. [ABSTRACT FROM AUTHOR]

The livelihoods of individuals in Ethiopia are severely affected by the livestock depredation from wild animals. The main objective of this study is to assess the incidences of livestock predation and pastoralist attitudes towards carnivore in the landscape of lions associated with Gambella National Park in the Abobo district of Western Ethiopia. The study was conducted between September 2017 and May 2018, utilizing a pretested semistructured questionnaire. Data analysis involved the utilization of multiple response analysis, Chi square test, Cramer's V and Cohen's Omega, Potential for Conflict Index2, and multinomial regression analysis. Anubis baboon (29.1%), leopard (20.5%), hyena (19.3%), lions (18.6%), and common fox (12.4%) were responsible for depriving cattle, sheep/goat, and fowls. The chi‐square test, along with Cramer's V and Cohen's Omega, indicated a relatively smaller effect size for the anubis baboon (ω = 0.220) and the common fox (ω = 0.164), implying a consistent trend of livestock depredation across villages. In contrast, medium effect size for leopards (ω = 0.466), hyenas (ω = 0.449), and lions (ω = 0.324) indicated that livestock depredation across villages was moderately inconsistent. In terms of preventive measures, barns and active guarding were reported options to protect their livestock, with a small effect size (ω = 0.257). Moreover, increased trend of livestock predation was perceived by informants with small effect size (ω = 0.227). Many respondents expressed a negative attitude (73.51%) towards wild animals than positive (8.60%) and neutral (17.90%) attitudes with a small effect size (ω = 0.181). The Potential for Conflict Index2 revealed a low consensus for acceptance of livestock depredation by different predators and skewed responses indicating the predominance of negative attitudes, regardless of livestock losses to predators. The regression model revealed that sociodemographic elements such as age, education, and occupation influence people's attitude. Hence, sustainable and culturally acceptable conservation measures are required to mitigate human‐wildlife​ conflict. [ABSTRACT FROM AUTHOR]

Background: Coxiella burnetii, the causative agent of Q fever, and Rift Valley fever virus are two under-researched zoonotic pathogens in Ethiopia. Potential outbreaks of these diseases, in light of the high dependency of nomadic pastoralists on their livestock, poses a risk to both human and animal health in addition to risking the pastoralists livelihoods. Our study aimed to determine the seroprevalence and associated risk factors for Q fever and Rift Valley fever in pastoral communities in the Afar region of north-eastern Ethiopia. Methodology/Principal findings: This cross-sectional study screened pastoralists (n = 323) and their livestock (n = 1377) for IgG antibodies to Coxiella burnetii and Rift Valley fever virus. A seroprevalence for Q fever of 25.0% (95%CI 18.6–32.6) was found in pastoralists and 34.3% (95%CI 27.9–41.3) in livestock overall; with 51.9% in goats (95%CI 44.9–58.8), 39.9% in sheep (95%CI 24.6–51.2), 16.3% in camels (95%CI 10.4–24.6) and 8.8% in cattle (95%CI 5.0–15.0). For Rift Valley fever the seroprevalence in pastoralists was 6.1% (95%CI 3.3–11.0) and 3.9% (95%CI 2.6–5.7) in livestock overall; cattle had the highest seroprevalence (8.3%, 95%CI 3.3–19.2), followed by goats (2.7%; 95%CI 1.4–5.1), sheep (2.5%; 95%CI 1.0–5.9) and camels (1.8%; 95%CI 0.4–6.9). Human Q fever seropositivity was found to be associated with goat abortions (OR = 2.11, 95%CI 1.18–3.78, p = 0.011), while Rift Valley fever seropositivity in livestock was found to be associated with cattle abortions (OR = 2.52, 95%CI 1.05–6.08, p = 0.039). Conclusions/Significance: This study provides evidence for a notable exposure to both Q fever and Rift Valley fever in pastoralists and livestock in Afar. The outbreak potential of these pathogens warrants ongoing integrated human and animal surveillance requiring close collaboration of the human and animal health sectors with community representatives following a One Health approach. Author summary: Q fever and Rift Valley fever are two diseases that can affect both humans and animals, causing illness and death. These two diseases can cause large-scale outbreaks, not only affecting the health of individuals and communities but also resulting in a substantial economic loss at the individual, community, regional and national levels through losses in livestock products. We conducted a sero-epidemiological study in the Afar region of north-eastern Ethiopia to determine the presence of these two diseases in nomadic pastoralists and their livestock. Our results indicate that 25.0% of the pastoralists and 34.3% of their animals have had previous exposure to Q fever and 6.1% of the pastoralists and 3.9% of their animals have been exposed to Rift Valley fever. Goats appear to be the most common livestock species associated with Q fever in this region, while cattle are the most common species associated with Rift Valley fever. Our findings highlight the importance of continuous surveillance; additionally, we would recommend the development of rapid response plans for potential outbreaks that integrate the human and animal health sectors. [ABSTRACT FROM AUTHOR]

Simple Summary: Getah virus (GETV) is an important zoonotic pathogen with an extensive host range, including horses, pigs, cattle, sheep, birds, and humans, resulting in substantial losses in the livestock and agricultural industries; as such, the prevalence and impact of GETV are significant concerns in China. In this study, the antigen domain of the E2 glycoprotein of GETV was expressed and purified in the Escherichia coli expression system, and an on-site immunochromatographic strip (ICS) was successfully developed to detect GETV antibodies in horses. The ICS has the advantages of being rapid, sensitive, specific, and is an effective method for the detection of GETV in horses that does not rely on special equipment or skilled personnel and can be used for the field diagnosis of GETV in horses. The prevalence and impact of Getah virus (GETV) are significant concerns in China. GETV can infect a wide range of animals, including horses, pigs, sheep, cattle, birds, and humans, resulting in substantial losses in the livestock and agricultural industries. GETV infection can cause the development of ulcers and inflammation in the mouth and gums of horses, which result in pain and discomfort and lead to symptoms such as reduced appetite, drooling, and difficulty chewing. As a result, there is a pressing need for efficient and rapid disease diagnosis methods. However, the currently available diagnostic methods have limitations in terms of operational time, equipment, and the experience of the individuals using them. In this study, a rapid, specific, and sensitive detection method was developed using a colloidal gold-based immunochromatographic strip (ICS) for the detection of antibodies against GETV in horses. To prepare the ICS, the antigen domain of the E2 glycoprotein of GETV was expressed using the Escherichia coli expression system after analysis with DNAstar v7.1 software. The nitrocellulose membrane was coated with rE2 protein or SPA to form the test line and control line, respectively. After optimizing the reaction conditions, the sensitivity, specificity, and repeatability of the strip were verified. The results showed that the test strip had a detection limit of up to 1:320 dilutions for GETV-positive serum, with no cross-reactivity observed with other equine-susceptible pathogens such as equine arteritis virus (EAV), equine herpesvirus-1 (EHV-I), equine infectious anemia virus (EIAV), equine influenza virus (EIV), African horse sickness virus (AHSV), and Japanese encephalitis virus (JEV). Furthermore, the ICS exhibited a concordance rate of 94.0% when testing 182 clinical serum samples compared to the virus neutralization test. Overall, this ICS diagnosis method will be an effective tool for the rapid detection of GETV in the field. [ABSTRACT FROM AUTHOR]

Haemonchus contortus (the barber's pole worm)—a highly pathogenic gastric nematode of ruminants—causes significant economic losses in the livestock industry worldwide. H. contortus has become a valuable model organism for both fundamental and applied research (e.g., drug and vaccine discovery) because of the availability of well-defined laboratory strains (e.g., MHco3(ISE).N1 in the UK and Haecon-5 in Australia) and genomic, transcriptomic and proteomic data sets. Many recent investigations have relied heavily on the use of the chromosome-contiguous genome of MHco3(ISE).N1 in the absence of a genome for Haecon-5. However, there has been no genetic comparison of these and other strains to date. Here, we assembled and characterised the mitochondrial genome (14.1 kb) of Haecon-5 and compared it with that of MHco3(ISE).N1 and two other strains (i.e., McMaster and NZ_Hco_NP) from Australasia. We detected 276 synonymous and 25 non-synonymous single nucleotide polymorphisms (SNPs) within Haecon-5. Between the Haecon-5 and MHco3(ISE).N1 strains, we recorded 345 SNPs, 31 of which were non-synonymous and linked to fixed amino acid differences in seven protein-coding genes (nad5, nad6, nad1, atp6, nad2, cytb and nad4) between these strains. Pronounced variation (344 and 435 SNPs) was seen between Haecon-5 and each of the other two strains from Australasia. The question remains as to what impact these mitogenomic mutations might have on the biology and physiology of H. contortus, which warrants exploration. The high degree of mitogenomic variability recorded here among these strains suggests that further work should be undertaken to assess the nature and extent of the nuclear genomic variation within H. contortus. [ABSTRACT FROM AUTHOR]

The orf virus (ORFV) poses a serious threat to the health of domestic small ruminants (i.e., sheep and goats) and humans on a global scale, causing around $150 million in annual losses to livestock industry. However, the host factors involved in ORFV infection and replication are still elusive. In this study, we compared the RNA-seq profiles of ORFV-infected or non-infected sheep testicular interstitial cells (STICs) and identified a novel host gene, potassium voltage-gated channel subfamily E member 4 (KCNE4), as a key host factor involved in the ORFV infection. Both RNA-seq data and RT-qPCR assay revealed a significant increase in the expression of KCNE4 in the infected STICs from 9 to 48 h post infection (hpi). On the other hand, the RT-qPCR assay detected a decrease in ORFV copy number in both the STICs transfected by KCNE4 siRNA and the KCNE4 knockout (KO) HeLa cells after the ORFV infection, together with a reduced fluorescence ratio of ORFV-GFP in the KO HeLa cells at 24 hpi, indicating KCNE4 to be critical for the ORFV infection. Furthermore, the attachment and internalization assays showed decreased ORFV attachment, internalization, replication, and release by the KO HeLa cells, demonstrating a potential inhibition of ORFV entry into the cells by KCNE4. Pretreatment with the KCNE4 inhibitors such as quinidine and fluoxetine significantly repressed the ORFV infection. All our findings reveal KCNE4 as a novel host regulator of the ORFV entry and replication, shedding new insight into the interactive mechanism of ORFV infection. The study also highlights the K+ channels as possible druggable targets to impede viral infection and disease. [ABSTRACT FROM AUTHOR]

Enterotoxigenic Escherichia coli (ETEC) are significant pathogen in both cattle and pigs, causing diarrhea in these animals and leading to economic losses in the livestock industry. Understanding the dissimilarity in genotype, antimicrobial resistance (AMR), and virulence between bovine and swine ETEC is crucial for development of targeted preventive and therapeutic approaches for livestock. However, a comprehensive study on this area remains lacking. Here, we performed whole-genome sequencingbased analyses of bovine (n = 554) and swine (n = 623) ETEC collected in the United States over a 53-year period. We identified distinct ETEC genotypes (fimH type, O antigen, H antigen, sequence type) in cattle and pigs. Furthermore, specific AMR and virulence profiles were associated with bovine and swine ETEC. Compared to swine ETEC, bovine ETEC were less diverse in genotypes and had a significantly (P < 0.001) lower number of AMR genes per isolate but higher co-occurrence of Shiga toxin and enterotoxin genes. Our results provide an overview of the key genomic differences between bovine and swine ETEC in the United States, which might be attributed to host adaptation and antibiotic usage practice. Ongoing surveillance and research are essential to monitor the genetic diversity and AMR patterns of ETEC in different host species. [ABSTRACT FROM AUTHOR]

Background and Aim: Theileria annulata infection in cattle causes major economic losses in livestock production in many Central Asian countries, including the southern region of Kazakhstan. This study aimed to obtain a recombinant T. annulata surface protein (TaSP) and to investigate its possible use as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of bovine theileriosis. Materials and Methods: Recombinant TaSP was obtained by cloning a polymorphic region of the TaSP gene, expressing it in Escherichia coli strain BL21, and purifying it by metal chelating chromatography. An indirect ELISA using recombinant TaSP as an antigen was developed and evaluated for the detection of T. annulata-specific antibodies in plasma samples from 69 cows polymerase chain reaction (PCR)-positive or PCR-negative for T. annulata and/or Theileria orientalis from southern Kazakhstan. Results: The obtained recombinant protein had a molecular weight of 32 kDa, and mass spectrometry analysis of the purified protein identified it as a fragment of the surface protein of T. annulata. Initial testing of 69 field plasma samples from cattle showed that the results of indirect ELISA using TaSP as an antigen agreed substantially with those of T. annulata PCR (κ: 0.78). The relative sensitivity and specificity of indirect ELISA were 88.7% and 100%, respectively, using PCR as a reference. There was no evidence of cross-reaction with T. orientalis. Conclusion: Initial results using recombinant TaSP as an antigen in indirect ELISA are promising and support the widespread use of this assay for routine diagnosis and T. annulata seroprevalence studies in cattle in Kazakhstan and possibly neighboring countries. [ABSTRACT FROM AUTHOR]

Bovine viral diarrhea virus (BVDV) is one of the most important etiological agents that causes serious economic losses to the global livestock industry. Vaccines usually provide limited efficacy against BVDV due to the emergence of mutant strains. Therefore, developing novel strategies to combat BVDV infection is urgently needed. Bergamottin (Berg), a natural furanocoumarin compound, possesses various pharmaceutical bioactivities, but its effect on BVDV infection remains unknown. The present study aimed to investigate the antiviral effect and underlying mechanism of Berg against BVDV infection. The results showed that Berg exhibited an inhibitory effect on BVDV replication in MDBK cells by disrupting the viral replication and release, rather than directly inactivating virus particles. Mechanistically, Berg inhibits BVDV replication by suppressing endoplasmic reticulum (ER) stress-mediated apoptosis via reducing reactive oxygen species (ROS) generation. Studies in vivo demonstrated that oral gavage of Berg at doses of 50 mg/kg and 75 mg/kg significantly reduced the viral load within the intestines and spleen in BVDV-challenged mice. Furthermore, histopathological damage and oxidative stress caused by BVDV were also mitigated with Berg treatment. Our data indicated that Berg suppressed BVDV propagation both in vitro and in vivo, suggesting it as a promising antiviral option against BVDV. [ABSTRACT FROM AUTHOR]

Mycoplasma bovis (M. bovis) is a significant bovine pathogen associated with various diseases, including bovine bronchopneumonia and mastitis resulting in substantial economic losses within the livestock industry. However, the development of effective control measures for M. bovis is hindered by a limited understanding of its virulence factors and pathogenesis. Nucleomodulins are newly identified secreted proteins of bacteria that internalize the host nuclei to regulate host cell gene expression and serve as critical virulence factors. Although recent reports have initiated exploration of mycoplasma nucleomodulins, the efficiency of conventional techniques for identification is very limited. Therefore, this study aimed to establish high-throughput methods to identify novel nucleomodulins of M. bovis. Using a direct biotinylation (DB) approach, a total of 289 proteins were identified including 66 high abundant proteins. In parallel, the use of proximity-based biotinylation (PBB), identified 28 proteins. Finally, seven nucleomodulins were verified to be nuclear by transfecting the bovine macrophage cell line BoMac with the plasmids encoding EGFP-fused proteins and observed with Opera Phenix, including the known nucleomodulin MbovP475 and six novel nucleomodulins. The novel nucleomodulins were four ribosomal proteins (MbovP599, MbovP678, MbovP710, and MbovP712), one transposase (MbovP790), and one conserved hypothetical protein (MbovP513). Among them, one unique nucleomodulin MbovP475 was identified with DB, two unique nucleomodulins (MbovP513 and MbovP710) with PBB, and four nucleomodulins by both. Overall, these findings established a foundation for further research on M. bovis nucleomodulin-host interactions for identification of new virulence factors. [ABSTRACT FROM AUTHOR]

Toxoplasma (T.) gondii and Neospora (N.) caninum are two significant abortifacient apicomplexan protozoa causing severe economic losses in livestock farming. This study was designed to investigate the serological status of T. gondii and N. caninum in sheep and goats across the Southeastern Anatolia Region of Türkiye, including some provinces bordering Syria and Iraq. Totally, 906 serum samples collected from female sheep (n=510) and goats (n=396) were investigated for specific antibodies against these protozoa by recombinant TgSAG2 and NcSAG1 protein-based indirect ELISA (iELISA). The individual seroprevalence of T. gondii and N. caninum in sheep was determined as 32.6 and 3.1%, respectively, while the seroprevalence of infection with both species was 33.1%. On the other hand, T. gondii and N. caninum seroprevalence in goats were 32.1 and 20.2%, respectively, while the total seroprevalence of infection with both species was 39.4%. The highest seroprevalence values for T. gondii (P<0.05) and N. caninum (P>0.05) were determined in sheep from Diyarbakır and Mardin provinces, respectively, while in goats, higher seroprevalence values for both protozoa were determined in Diyarbakır province compared to other provinces (P<0.05). Statistical analyses revealed that seroprevalence values varying between provinces were significant (P<0.05), except for ovine neosporosis (P>0.05) and when seroprevalence values between animal species were compared, caprine neosporosis was statistically significantly higher (P<0.05). The fact that 35.9% of the small ruminants enrolled in the study had been exposed to abortive apicomplexan protozoa during their lives is a serious concern, given the zoonotic aspects of these protozoa and the economic damage they can cause. [ABSTRACT FROM AUTHOR]

Sheeppox virus (SPPV), goatpox virus (GTPV), and lumpy skin disease virus (LSDV) are the three members of the genus Capripoxvirus within the Poxviridae family and are the etiologic agents of sheeppox (SPP), goatpox (GTP), and lumpy skin disease (LSD), respectively. LSD, GTP, and SPP are endemic in Africa and Asia, causing severe disease outbreaks with significant economic losses in livestock. Incursions of SPP and LSD have occurred in Europe. Vaccination with live attenuated homologous and heterologous viruses are routinely implemented to control these diseases. Using the gold standard virus neutralization test, we studied the ability of homologous and heterologous sera to neutralize the SPPV and LSDV. We found that LSD and SPP sera effectively neutralize their homologous viruses, and GTP sera can neutralize SPPV. However, while LSD sera effectively neutralizes SPPV, SPP and GTP sera cannot neutralize the LSDV to the same extent. We discuss the implications of these observations in disease assay methodology and heterologous vaccine efficacy. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease virus (FMDV) is a highly contagious virus that affects cloven-hoofed animals and causes severe economic losses in the livestock industry. Given that this high-risk pathogen has to be handled in a biosafety level (BSL)-3 facility for safety reasons and the limited availability of BSL-3 laboratories, experiments on FMDV call for more attention. Therefore, we aimed to develop an FMDV experimental model that can be handled in BSL-2 laboratories. The NanoBiT luciferase (Nano-luc) assay is a well-known assay for studying protein–protein interactions. To apply the NanoBiT split luciferase assay to the diagnosis and evaluation of FMD, we developed an inactivated HiBiT-tagged Asia1 Shamir FMDV (AS-HiBiT), a recombinant Asia1 shamir FMDV with HiBiT attached to the VP1 region of Asia1 shamir FMDV. In addition, we established LgBiT-expressing LF-BK cell lines, termed LgBit-LF-BK cells. It was confirmed that inactivated AS-HiBiT infected LgBiT-LF-BK cells and produced a luminescence signal by binding to the intracellular LgBiT of LgBiT-LF-BK cells. In addition, the luminescence signal became stronger as the number of LgBiT-LF-BK cells increased or the concentration of inactivated AS-HiBiT increased. Moreover, we confirmed that inactivated AS-HiBiT can detect seroconversion in sera positive for FMDV-neutralizing antibodies. This NanoBiT split luciferase assay system can be used for the diagnosis and evaluation of FMD and expanded to FMD-like virus models to facilitate the evaluation of FMDV vaccines and antibodies. [ABSTRACT FROM AUTHOR]

This study was designed to provide data on the incidence rates of congenital anomalies and their distribution by organs and systems in calves, lambs and kids brought to Van Yuzuncu Yil University, Faculty of Vetrinary Medicine, Surgery Clinic between 2017-2023. The animal material of the study consisted of 1104 ruminants, including 886 calves, 183 lambs and 35 kids, aged 0-6 months. In addition to clinical examination, anomalies were diagnosed by using direct and indirect radiographic examinations when necessary. In some cases, a definitive diagnosis was made by performing experimental laparotomy based on clinical and radiological examinations. 341 out of 1104 ruminants, including 234 calves, 96 lambs and 11 kids, suffered from congenital anomalies. Of the ruminants with congenital anomalies, 208 (61.00%) were male, 132 (38.70%) female and 1 (0.30%) hermaphrodite. Abdominal wall anomalies were determined with the highest number of 112 (32.9%) cases. These were followed by musculoskeletal system anomalies in 83 (24.4%) cases and gastrointestinal system anomalies in 56 (16.5%) cases. Head region anomalies occurred in 41 cases (12.1%), urinary system anomalies in 23 cases (6.8%) and CNS anomalies in 9 cases (2.6%). Multiple anomalies were recorded in 16 (4.7%) cases. In conclusion, congenital anomalies are frequently seen as pathologies in ruminants in our country, especially in our region. Avoiding inbreeding, making artificial insemination widespread, improving the care and nutrition of the dam during pregnancy, correcting adverse environmental conditions and eliminating stress factors, avoiding over-the-counter medication misuse during pregnancy and most importantly informing field veterinarians and animal owners about the subject will contribute to the prevention of congenital malformations and losses in livestock. [ABSTRACT FROM AUTHOR]

Background: Contagious and economically devastating, peste des petits ruminants (PPR) is a viral disease affecting goats and sheep, causing significant losses in livestock productivity and posing a threat to food security and rural livelihoods worldwide. Objectives: This study was conducted to assess the status of goat farmer's knowledge, attitude and practice (KAP) about PPR disease at Sylhet district of Bangladesh. Methods: A comprehensive cross‐sectional survey, conducted over 11 weeks, targeted 130 goat owners. Following a pilot study with 20 farmers, a set of 17 validated questions on PPR KAP was validated. Data collection was performed through face‐to‐face interviews by a trained team using KOBO Toolbox, with interpretation of responses based on established thresholds for knowledge (>65%), attitude (>75%) and practice (>70%). Results: Overall, 67.7% of participants demonstrated a good level of knowledge about PPR disease. Males exhibited 1.42 times higher odds of knowledge compared to females (odds ratio = 1.42). The middle age group (31–45 years) showed significantly higher knowledge levels (83.3%, p < 0.001). Within this age group, positive practice levels were also notably higher (54.8%). Those with a higher secondary education background exhibited the most positive practice levels (>80%). Participants whose additional income came from non‐governmental organization employment showed a higher positive practice level (71.4%), 1.46 times higher than other income sources. Conclusion: Strategic interventions should prioritize female farmers, educational empowerment and collaboration with non‐governmental organizations to bolster livestock health and rural livelihoods in Bangladesh as part of national PPR control strategy to fulfil the goals of Office International des Epizooties/World Organization for Animal Health (OIE/WOAH) and Food and Agriculture Organization (FAO) PPR eradication by 2030. [ABSTRACT FROM AUTHOR]

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This study investigated the dynamics of perceived livestock depredation over the last 2 years (from 2021 to 2022) in villages close to Mkomazi National Park in Tanzania. The findings revealed that sheep were the most frequently affected livestock (57.1%), followed by goats (36.7%) and cattle (6.1%). Attacks primarily occurred at home (59.2%) and during the night (65.3%), with a higher prevalence during the dry season (75.5%). The majority of respondents (81.6%) perceived a decrease in attacks. Logistic regression indicated village proximity to the park as a significant predictor in explaining the reported incidents related with livestock depredation, with closer villages experiencing higher risks. Gender, age, education level, and resident status showed no significant correlation with the reported livestock depredation incidents. Spotted hyena sightings varied significantly based on village proximity to the park (p = 0.039), while other factors showed no significant differences. Mitigation measures against livestock depredation included guarding livestock (28.3%), employing noise and scaring tactics (42.2%), and utilizing domestic dogs (29.4%). These findings offer valuable insights into human-wildlife conflicts and inform targeted interventions for mitigating livestock losses not only in the surveyed villages but also in other areas globally facing similar challenges. [ABSTRACT FROM AUTHOR]

One of the major management problems in and around protected areas is the intensifying conflict between local people and wildlife, especially large carnivores. Livestock depredation and human fatalities caused by attacks of carnivores are found to be serious obstacles in conflict management. This study aims to explore the patterns, costs, causes and perceptions of human large carnivore conflict in the Nawalpur area of Chitwan National Park. The patterns of livestock loss and human casualties due to large carnivores (Panthera tigris and Panthera pardus) were analysed using the secondary data reported to Chitwan National Park from 2001 to 2019. To understand the people's perception towards carnivores and wildlife conservation, 150 victim respondents were asked, with one household selected from each grid. During 19-year study period, a total of 521 incidents caused by large carnivores were reported, which included 33 human casualties and 488 livestock depredations. Tiger was responsible for the maximum conflict incidents in Nawalpur. The total relief provided for human deaths and injuries was US$17524.41, whereas US$13702.18 was used to compensate for livestock depredation in the Nawalpur area by authorities of the Chitwan National Park. More than 64% of the respondents liked the presence of carnivores in their area and had a positive attitude toward the conservation of large carnivores, even though the carnivores were responsible for livestock depredation and human injury and death. People's satisfaction with the relief scheme provided by the government depended on ethnicity, gender, age class, occupation, education, insurance, and livestock ownership, but they believed the scheme was not effective. Hence, an effective relief scheme and awareness about the process of relief funding should be conducted in conflict areas. Conducting awareness programs for local communities about large carnivores, their behavior, and preparing predator-proof corrals would be helpful in minimizing conflict in the study area. [ABSTRACT FROM AUTHOR]

Capripoxviruses are the causative agents of sheeppox, goatpox, and lumpy skin disease (LSD) in cattle, which cause economic losses to the livestock industry in Africa and Asia. Capripoxviruses are currently controlled using several live attenuated vaccines. It was previously demonstrated that a lumpy skin disease virus (LSDV) field isolate from Warmbaths (WB) South Africa, ORF 005 (IL-10) gene-deleted virus (LSDV WB005KO), was able to protect sheep and goats against sheeppox and goatpox. Subsequently, genes encoding the protective antigens for peste des petits ruminants (PPR) and Rift Valley fever (RVF) viruses have been inserted in the LSDV WB005KO construct in three different antigen forms (native, secreted, and fusion). These three multivalent vaccine candidates were evaluated for protection against PPR using a single immunization of 104 TCID50 in sheep. The vaccine candidates with the native and secreted antigens protected sheep against PPR clinical disease and decreased viral shedding, as detected using real-time RT-PCR in oral and nasal swabs. An anamnestic antibody response, measured using PPR virus-neutralizing antibody response production, was observed in sheep following infection. The vaccine candidates with the antigens expressed in their native form were evaluated for protection against RVF using a single immunization with doses of 104 or 105 TCID50 in sheep and goats. Following RVF virus infection, sheep and goats were protected against clinical disease and no viremia was detected in serum compared to control animals, where viremia was detected one day following infection. Sheep and goats developed RVFV-neutralizing antibodies prior to infection, and the antibody responses increased following infection. These results demonstrate that an LSD virus-vectored vaccine candidate can be used in sheep and goats to protect against multiple viral infections. [ABSTRACT FROM AUTHOR]

Toxoplasmosis is a cosmopolitan zoonotic infection that has significant effects on public health and causes economic losses in the livestock industry. The current study was designed to detect the Toxoplasma parasite in sheep blood samples and tissue samples of slaughtered sheep at the Sulaimani abattoir using molecular technique. A total of 300 peripheral sheep blood samples were randomly collected from 20 small ruminant flocks at 4 locations in the Sulaymaniyah province, northern Iraq. Also, 150 meat samples from thigh muscle, heart, and diaphragm were collected from slaughtered sheep. All collected blood samples were subjected to polymerase chain reaction (PCR) amplification to confirm Toxoplasma infection; in addition, meat samples were also analyzed for Toxoplasma by PCR following the digestion process. Of the 300 amplified blood samples, 94 were considered positive for Toxoplasma gondii, with a prevalence rate of 31.3%. The overall prevalence of Toxoplasma among meat samples was 34%. The diaphragm reported a higher infection rate (46%) than the heart (32%), while the femoral muscle reported an infection rate of 24%. Aged animals (older than 24 months) presented a higher infection rate (32.8%) than younger animals (28.9%). Contact with or consumption of uncooked meat from infected sheep increases the chance of parasite transmission to humans. [ABSTRACT FROM AUTHOR]

The article is about calving in cattle and its stages. It is mentioned that the transition is a period of approximately 42 days that covers three weeks before and three weeks after calving. Calving is a key stage in the cow's transition, as it conditions her productive and reproductive life. Additionally, it is mentioned that dystocia, or difficulty in calving, causes economic losses in livestock due to increased calf mortality, calf morbidity in the first 30 days of life, trauma to the birth canal in the mother, decreased milk production, and increased risk of involuntary culling. The premonitory signs of calving and its stages are also mentioned. The cervix actively participates in its dilation before calving in cows. Uterine contractions begin at the end of gestation and gradually increase until the moment of calving. During the cervical dilation phase, the fetus acquires righting reflexes and extends its carpal joints. The expulsion of the fetus occurs through uterine and abdominal contractions. After calving, the expulsion of the placenta occurs. It is recommended to perform obstetric examination before and after calving to detect possible complications. The article discusses the importance of separating the calf from its mother immediately after calving to reduce the risk of infections and maternal stress. It is mentioned that contact between the mother and the calf improves antibody absorption in the newborn calf. The use of a "cuddle box" is recommended to maintain the mother-calf relationship and reduce the risk of infection for the calf. The importance of disinfecting the calf's umbilicus and applying navel dips is also mentioned. The article highlights the importance of proper calving management for the cow's future productivity and reproductive success. [Extracted from the article]

Human-wildlife conflict (HWC) is a pressing issue worldwide but varies by species over time and place. One of the most prevalent forms of HWC in the mid-hills of Nepal is human-common-leopard conflict (HLC). Leopard attacks, especially in forested areas, can severely impact villagers and their livestock. Information on HLC in the Gorkha district was scarce, thus making it an ideal location to identify high-risk zones and landscape variables associated with such events. Registered cases were collected and reviewed from the Division Forest Office (DFO) during 2019-2021. Claims from DFO records were confirmed with herders and villagers via eight focus group discussions. To enhance modeling success, researchers identified a total of 163 leopard attack locations on livestock, ensuring a minimum distance of at least 100 meters between locations. Using maximum entropy (MaxEnt) and considering 13 environmental variables, we mapped common leopard attack risk zones. True Skill Statistics (TSS) and area under receiver-operator curve (AUC) were used to evaluate and validate the Output. Furthermore, 10 replications, 1,000 maximum iterations, and 1000 background points were employed during modeling. The average AUC value for the model, which was 0.726 ± 0.021, revealed good accuracy. The model performed well, as indicated by a TSS value of 0.61 ± 0.03. Of the total research area (27.92 km2), about 74% was designated as a low-risk area, 19% as a medium-risk area, and 7% as a high-risk area. Of the 13 environmental variables, distance to water (25.2%) was the most significant predictor of risk, followed by distance to road (16.2%) and elevation (10.7%). According to response curves, the risk of common leopard is highest in the areas between 1.5 to 2 km distances from the water sources, followed by the closest distance from a road and an elevation of 700 to 800 m. Results suggest that managers and local governments should employ intervention strategies immediately to safeguard rural livelihoods in high-risk areas. Improvements include better design of livestock corrals, insurance, and total compensation of livestock losses. Settlements near roads and water sources should improve the design and construction of pens and cages to prevent livestock loss. More studies on the characteristics of victims are suggested to enhance understanding of common leopard attacks, in addition to landscape variables. Such information can be helpful in formulating the best management practices. [ABSTRACT FROM AUTHOR]

Dicrocoelium lancet flukes cause significant production loss in ruminant livestock. Although co-infection with multiple Dicrocoelium species within a host is common, techniques for studying the composition of these complex parasite communities are lacking. The pathogenicity, epidemiology, and therapeutic susceptibility of different helminth species vary, and little is known about the interactions that take place between co-infecting species and their hosts. Here, we describe the first applicationof metabarcoding deep amplicon sequencing method to studythe Dicrocoelium species in sheep and goats. First, rDNA ITS-2 sequences of four Dicrocoelium species (Dicrocoelium dendriticum, Dicrocoelium hospes, Dicrocoelium orientalis, and Dicrocoelium chinensis) were extracted from the NCBI public database. Phylogenetic analysis revealed separate clades of Dicrocoelium species; hence, molecular differentiation between each species is possible in co-infections. Second, 202 flukes belonging to seventeen host populations (morphologically verified as belonging to the Dicrocoelium genus) were evaluated to determine the deep amplicon sequencing read threshold of an individual fluke for each of the four species. The accuracy of the method in proportional quantification of samples collected from single hosts was further assessed. Overall, 198 (98.01%) flukes were confirmed as D. dendriticum and 1.98% produced no reads. The comparison of genetic distances between rDNA ITS-2 revealed 86% to 98% identity between the Dicrocoelium species. Phylogenetic analysis demonstrated a distinct clustering of species, apart from D. orientalis and D. chinensis, which sit very close to each other in a single large clade whereas D. hospes and D. dendriticum are separated into their own clade. In conclusion each sample was identified as D. dendriticum based on the proportion of MiSeq reads and validated the presence of this group of parasites in the Gilgit Baltistan and Khyber Pakhtunkhwa provinces of Pakistan. The metabarcoding deep amplicon sequencing technology and bioinformatics pathway have several potential applications, including species interactions during co-infections, identifying the host and geographical distribution of Dicrocoelium in livestock, drug therapy response evaluation and understanding of the emergence and spread of drug resistance. [ABSTRACT FROM AUTHOR]

Bovine Viral Diarrhea Virus (BVDV) and Infectious Bovine Rhinotracheitis Virus (IBRV) are the two most prevalent infectious diseases in cattle. They both can cause persistent infection and immunosuppression, resulting in significant economic losses in the livestock industry. Therefore, rapid detection of early BVDV and IBRV infections is crucial. In this study, a method for the rapid detection of BVDV and IBRV was established by using recombinase polymerase amplification (RPA) combined with lateral flow device (LFD). By optimizing the temperature and time conditions of the RPA reaction, the sensitivity, specificity, and clinical performance were evaluated. The results indicated that the RPA reaction could be completed at 40◦C within 25min. The LOD for BVDV and IBRV by RPA-LFD were 5.1 × 10¹ copies/μL and 6.65 × 10¹ copies/μL, respectively, with no cross-reactivity observed with other viruses such as CSFV, BRSV, BPIV3, BRV, and BCoV. Testing of 32 clinical samples showed consistent results between RPA-LFD and qPCR. The RPA-LFD method established in this study can be used for the rapid clinical detection of BVDV and IBRV, which providing a rapid and convenient molecular biology approach for on-site rapid detection and epidemiological investigations. Simultaneously, it offers technical support for the prevention and control of these viruses. [ABSTRACT FROM AUTHOR]

Coxiella burnetii is a highly infectious bacterium responsible for causing Q fever, an emerging public health problem of global concern and the cause of severe economic losses in livestock farming. Many species can be asymptomatic carriers and shed this bacterium in various secretions and excreta. Humans can acquire the infection mainly through environmental contamination due to bacterial shedding of infected animals, and through tick-borne or foodborne transmissions. If this agent is aerosolized, it is considered a potential biological weapon. Advanced molecular and serological diagnostic techniques for Q fever over the last decade have made it possible to detect clinical cases and carry out population screening effectively. There needs to be more research on C. burnetii control in wildlife despite the increasing evidence that wildlife is a source of C. burnetii for both domestic animals and humans. Birds can transmit this pathogen directly or indirectly to other animals or humans, but data about the spreading of C. burnetii in avian populations still need to be available. The present work aimed to revise the literature about the involvement of wild birds in the epidemiology of Q fever in Europe. Reports of this pathogenic agent in wild birds in European countries since 2007 were considered. [ABSTRACT FROM AUTHOR]

Introduction: Goatpox, a severe infectious disease caused by goatpox virus (GTPV), leads to enormous economic losses in the livestock industry. Traditional live attenuated vaccines cause serious side effects and exist a risk of dispersal. Therefore, it is urgent to develop efficient and safer vaccines to prevent and control of GTPV. Methods: In the present study, we are aimed to design a multi-epitope subunit vaccine against GTPV using an immunoinformatics approach. Various immunodominant cytotoxic T lymphocytes (CTL) epitopes, helper T lymphocytes (HTL) epitopes, and B-cell epitopes from P32, L1R, and 095 proteins of GTPV were screened and liked by the AAY, GPGPG, and KK connectors, respectively. Furthermore, an adjuvant b-defensin was attached to the vaccine's N-terminal using the EAAAK linker to enhance immunogenicity. Results: The constructed vaccine was soluble, non-allergenic and non-toxic and exhibited high levels of antigenicity and immunogenicity. The vaccine's 3D structure was subsequently predicted, refined and validated, resulting in an optimized model with a Z-value of -3.4. Molecular docking results demonstrated that the vaccine had strong binding affinity with TLR2(-27.25 kcal/mol), TLR3(-39.84 kcal/mol), and TLR4(-59.42 kcal/mol). Molecular dynamics simulation results indicated that docked vaccine-TLR complexes were stable. Immune simulation analysis suggested that the vaccine can induce remarkable increase in antibody titers of IgG and IgM, higher levels of IFN-g and IL-2. Conclusion: The designed GTPV multi-epitope vaccine is structurally stable and can induce robust humoral and cellular immune responses, which may be a promising vaccine candidate against GTPV. [ABSTRACT FROM AUTHOR]

Background: Contagious bovine pleuropneumonia [CBPP] is a transboundary animal disease of cattle caused by Mycoplasma mycoides subsp. mycoides [Mmm]. CBPP causes severe economic losses to livestock producers in sub-Saharan Africa mainly due to high mortality, morbidity, reduction in productivity as well as livestock trade restrictions. This study aimed at determining seroprevalence of Mmm in cattle from Karamoja region, north-eastern Uganda; data that are required to design and implement risk based CBPP control program. Methods: We randomly collected blood samples from 2,300 cattle spread across Karamoja region. Serum was extracted and screened for antibodies against Mycoplasma mycoides subsp. mycoides [Mmm] using the competitive enzyme linked immunosorbent assay [cELISA]. Results: A quarter [25.4%; 95% CI: 23.7–27.3] of the screened cattle [n = 2,300] were sero-positive for Mmm. Amudat and Kaabong districts recorded the lowest [12.3%] and highest [30.7%] Mmm seroprevalence respectively. Increasing age, overnight stay in cattle kraals and location [certain districts, villages, herds and sub counties] of the cattle herds, the factors that promote animal commingling, were the most significant risk factors of seroconversion with Mmm. Conclusion: Results from this study indicated a higher seroprevalence of Mmm in Karamoja region cattle herds. This could be due to the increased frequency of CBPP outbreaks in recent years. To be effective, CBPP vaccination programs should target high risk herds along the international borders and other hotspot areas [e.g., parishes or sub counties] where cattle commingling is high. [ABSTRACT FROM AUTHOR]