The risk of pathogen transmission continues to increase significantly in the presence of tick vectors due to the trade of livestock across countries. In Ghana, there is a lack of data on the incidence of tick-borne pathogens that are of zoonotic and veterinary importance. This study, therefore, aimed to determine the prevalence of such pathogens in livestock using molecular approaches. A total of 276 dry blood spots were collected from cattle (100), sheep (95) and goats (81) in the Kassena-Nankana Districts. The samples were analyzed using Polymerase Chain Reaction (qPCR) and conventional assays and Sanger sequencing that targeted pathogens including Rickettsia, Coxiella, Babesia, Theileria, Ehrlichia and Anaplasma. An overall prevalence of 36.96% was recorded from the livestock DBS, with mixed infections seen in 7.97% samples. Furthermore, the prevalence of infections in livestock was recorded to be 19.21% in sheep, 14.13% in cattle, and 3.62% in goats. The pathogens identified were Rickettsia spp. (3.26%), Babesia sp. Lintan (8.70%), Theileria orientalis (2.17%), Theileria parva (0.36%), Anaplasma capra (18.48%), Anaplasma phagocytophilum (1.81%), Anaplasma marginale (3.26%) and Anaplasma ovis (7.25%). This study reports the first molecular identification of the above-mentioned pathogens in livestock in Ghana and highlights the use of dry blood spots in resource-limited settings. In addition, this research provides an update on tick-borne pathogens in Ghana, suggesting risks to livestock production and human health. Further studies will be essential to establish the distribution and epidemiology of these pathogens in Ghana.
Doxycycline hydrochloride and florfenicol combination (DoxHcl&FF) is an effective treatment for respiratory diseases. In the study, our objective was to evaluate the activity of DoxHcl&FF against Actinobacillus pleuropneumoniae (APP) in porcine pulmonary epithelial lining fluid (PELF) and the optimal dosage scheme to avoid the development of resistance. The DoxHcl&FF was administered intramuscularly (IM) at 20 mg/kg, and the PELF was collected at different time points. The minimum inhibitory concentration (MIC) and time-mortality curves were also included in the study. Based on the sigmoid Emax equation and dose equations, the study integrated the in vivo pharmacokinetic data of infected pigs and ex vivo pharmacodynamic data to obtain the area under concentration time curve (AUC0-24h)/MIC values in PELF and achieve bacteriostatic activity, bactericidal activity and the virtual eradication of bacteria. The study showed that the combination of DoxHcl and FF caused no significant changes in PK parameters. The peak concentration (Cmax) of FF in healthy and diseased pigs was 8.87 ± 0.08 μg/mL and 8.67 ± 0.07 μg/mL, the AUC0-24h were 172.75 ± 2.52 h·μg/mL and 180.22 ± 3.13 h·μg/mL, the Cmax of DoxHcl was 7.91 ± 0.09 μg/mL and 7.99 ± 0.05 μg/mL, and the AUC0-24h was 129.96 ± 3.70 h·μg/mL and 169.82 ± 4.38 h·μg/mL. DoxHcl&FF showed strong concentration-dependent tendencies. The bacteriostatic, bactericidal, and elimination activity were calculated as 5.61, 18.83 and 32.68 h, and the doses were 1.37 (bacteriostatic), 4.59 (bactericidal) and 7.99 (elimination) mg/kg. These findings indicated that the calculated recommended dose could assist in achieving more precise administration, increasing the effectiveness of DoxHcl&FF treatment for APP infections.
Inappropriate use of antibiotics is globally creating public health hazards associated with antibiotic resistance. Bacteria often acquire antibiotic resistance by altering their genes through mutation or acquisition of plasmid-encoding resistance genes. To treat drug-resistant strains of bacteria, the recently developed CRISPR-Cas9 system might be an alternative molecular tool to conventional antibiotics. It disables antibiotic-resistance genes (plasmids) or deactivates bacterial virulence factors and sensitizes drug-resistant bacteria through site-specific cleavage of crucial domains of their genome. This molecular tool uses phages as vehicles for CRISPR-cas9 delivery into bacteria. Since phages are species-specific and natural predators of bacteria, they are capable of easily injecting their DNA to target bacteria. The CRISPR system is packaged into phagemid vectors, in such a way that the bacteria containing the antibiotic-resistance plasmid sequence or that containing specific DNA sequences were made to be targeted. Upon CRISPR delivery, Cas9 is programmed to recognize target sequences through the guide RNA thereby causing double-strand cleavage of targeted bacterial DNA or loss of drug resistance plasmid, which results in cell death. Remarkably, the safety and efficacy of this newly developed biotechnology tool and the biocontrol product need to be further refined for its usage in clinical translation.
Mycobacterium tuberculosis (M. tuberculosis) can replicate in the macrophage by interfering with many host protein functions. While it is far from known these host proteins for controlling M. tuberculosis infection. Herein, we infected macrophages including THP-1 and Raw264.7 cells with M. tuberculosis and identified the differentially expressed genes (DEGs) in the interferon signaling pathway. Among them, 2′-5′ oligoadenylate synthetase-like (OASL) underwent the greatest upregulation in M. tuberculosis-infected macrophages. Knockdown of the expression of OASL attenuated M. tuberculosis survival in macrophages. Further, bioinformatics analysis revealed the potential interaction axis of OASL-TAB3- Rv0127, which was further validated by the yeast-two-hybrid (Y2H) assay and Co-IP. This interaction axis might regulate the M. tuberculosis survival and proliferation in macrophages. The study reveals a possible role of OASL during M. tuberculosis infection as a target to control its propagation.
The cortex of the limb bones of chinchillas is very thin and brittle, so it is prone to fractures of the limb bones, among which fractures of the tibia, radius and ulna are the most frequent types. When a chinchilla has a closed fracture, it can be immobilized with a splint, cast, or bandage. If the broken end of the fracture pierces the skin, it is best to choose internal fixation or external fixation brackets for treatment. In this report, a 0.661 kg, 2-year-old male uncastrated chinchilla was presented to the Veterinary Teaching Hospital of Huazhong Agricultural University due to an old fracture of the right forearm. With the consent of the owner, we decided to use a 25-gauge needle as an IM pin to fix the fracture. Ten days after surgery, the wound had healed well, and the limb could support body weight, but the palm did not show a grasping position. Twenty four days after the operation, the affected limb had not regained the ability to grasp. The X-ray showed a slight rotation of the IM pin and good callus growth in the ulna, but not in the radius. One month after the operation, it was found that the function of the affected limb of the chinchilla was normal and the grasping ability was restored through follow-up consultation and the return visit.
Mosquito-borne infections are of global health concern because of their rapid spread and upsurge, which creates a risk for coinfections. chikungunya virus (CHIKV), an arbovirus disease transmitted by Aedes aegypti or A. albopictus, and malaria, a parasitic disease transmitted by Anopheles gambiae, are prevalent in Nigeria and neighbouring countries, but their burden and possible coinfections are poorly understood. In this study, we investigated the antibody seropositivity and endemicity of chikungunya and Zika viruses (ZIKV) in three regions of Nigeria. A cross-sectional sero-survey was conducted on 871 participants. Samples were collected from outpatients by simple random sampling. Analyses of the samples were performed using recomLine Tropical Fever for the presence of antibody serological marker IgG immunoblot with CHIKV VLP (virus like particle), ZIKV NS1 and ZIKV Equad according to manufacturers’ instructions and malaria RDT for malaria parasite. There was a significantly higher antibody seropositivity against CHIKV in the central region than in the northern and southern regions (69.5%, 291/419), while ZIKV-seropositivity (22.4%, 34/152) and CHIKV-ZIKV co-circulating antibody seropositivity (17.8%, 27/152) were notably higher in the southern region than in the central and northern regions. This investigation revealed an unexpectedly high antibody seropositivity and concealed endemicity of CHIKV and ZIKV in three Nigerian regions. The seropositivity of detectable antibodies differed among the three geographical locations.
Bovine tuberculosis (bTB) is a chronic zoonotic disease that is endemic in China. Current in-vitro tests for bTB are mainly based on blood assays. Collection of samples results in some stress to the sampled cattle and associated economic losses for the herd owner. This study was designed to investigate the relationship between milk and serum antibody tests for bTB in dairy cows using 85 cows with milk and corresponding blood samples. Totally 4,395 milk samples were used to assesse the apparent (test) prevalence and incidence of bTB using the milk antibody ELISA. The association between levels of bTB milk antibody and milk quality was also evaluated. Milk and serum antibody tests showed a good correlation with a 87.5% (95% CI: 61.7%, 98.4) positive agreement and 98.7% (95% CI: 95.4, 99.8) negative agreement. The animal level lactoprevalence ranged from 0.3% (95% CI: 0, 1.2) to 33.3% (95% CI: 26.6, 40.6) in different farms and the incidence rate ranged from 0 head/cow-month (95% CI: 0, 0.02) to 0.04 head/cow-month (95% CI: 0.02, 0.07). Twenty percent of sampled farms met the criteria for bTB control in China. The prevalence on large-scale farms was lower (p < 0.001) than on small farms. The bTB milk antibody levels had a negative correlation with milk yield and a positive correlation with somatic cell count (SCC), milk protein percentage (MPP) and percentage of total solids (TS). According to this research, milk ELISA could be used as a supplement of blood samples to assist in the surveillance for bTB and for alerting control and eradication of bTB.
Taenia solium taeniasis and cysticercosis are neglected zoonotic diseases that affect human health and economies of developing countries. In this work, we formulate and analyze deterministic and continuous time Markov chain (CTMC) stochastic models to determine parameters that drive Taenia solium taeniasis and cysticercosis and the likelihood of their extinction. The basic reproduction number $R_0$ is computed by the next generation matrix approach, sensitivity index of each parameter in $R_0$ is derived by the normalized forward sensitivity index and the likelihood of diseases’ extinction is computed by the multitype branching process. The analysis shows that humans with Taenia solium taeniasis, infectious pork and Taenia solium eggs in the environment play an important role in the transmission of Taenia solium taeniasis and cysticercosis, and the model exhibits forward bifurcation at $R_0=1$. This implies that $R_0<1$ is a sufficient condition to eliminate Taenia solium taeniasis and cysticercosis. For CTMC model, analysis shows that the probability of Taenia solium taeniasis and cysticercosis extinction is high if the diseases emerge from humans with Taenia solium cysticercosis and there is an outbreak if the diseases emerge from either humans with Taenia solium taeniasis or infectious pork or Taenia solium eggs in the environment. To control Taenia solium taeniasis and cysticercosis, the intervention strategies should focus on improving hygiene and sanitation for reducing shedding rate of Taenia solium eggs in the environment, inspection of pork for reducing the rate of acquiring Taenia solium taeniasis and spraying of insecticides for killing Taenia solium eggs in the environment.
Lamb morbidity and mortality cause significant loss in a smallholder production system. A longitudinal prospective survey was conducted on 408 randomly selected farmers located in six purposefully selected kebeles in Gewata woreda from June 2020 to July 2021 to determine the incidence of morbidity and mortality in lambs and to identify risk factors. For this purpose, 408 lambs from the Gewata district’s mixed crop-livestock production system were examined every day from birth to three months of age. The data was analyzed using the Kaplan-Meier (K-M) method, the log-rank test, and Cox proportional hazards regression. Results showed that the cumulative incidence of all-cause of morbidity and mortality at the end of three month was 12.86% (95% CI: 10.26–16.13%), and 6.86% (95% CI: 5.03–9.35%), respectively. Diarrhea was the leading cause of morbidity and mortality, accounting for 49.33 and 50% morbidity and mortality, respectively. Pneumonia was the second most frequent cause of morbidity and death. In the K-M hazard analysis, the greatest risk of lamb morbidity and mortality was observed during the first month of life, and then the risk decreased significantly as the lamb grew. Of the 17 potential risk factors studied, the multivariable Cox proportional hazards regression model showed that lamb sex, birth weight, umbilical care, time and method of colostrum feeding were the five predictors that were significantly associated with a higher risk of morbidity, whereas birth weight, lambing difficulty, dams’ parity, method and time of colostrum feeding were the five predictors that were significantly associated with a higher risk of mortality. Moreover, a higher risk of morbidity was observed in lambs with methods of colostrum feeding after birth (HR = 3.158; p = 0.000) and with variations in birth weight (HR = 1.418; p = 0.003). Similarly, the mortality risk was 4.926 (p = 0.047), 4.023 (p = 0.012), and 3.206 (p = 0.000) times higher in lambs with lambing difficulties, at the time of colostrum feeding, and by the method of colostrum feeding, respectively. According to this research, lamb morbidity and mortality rates in the study area are significantly high which has a great impact on the replacement stock and production. Hence, awareness should be created among farmers concerning improved lamb management practices.
The spike protein (S) of SARS-CoV-2 is responsible for viral attachment and entry, thus a major factor for host susceptibility, tissue tropism, virulence and pathogenicity. The S is divided with S1 and S2 region, and the S1 contains the receptor-binding domain (RBD), while the S2 contains the hydrophobic fusion domain for the entry into the host cell. Numerous host proteases have been implicated in the activation of SARS-CoV-2 S through various cleavage sites. In this article, we review host proteases including furin, trypsin, transmembrane protease serine 2 (TMPRSS2) and cathepsins in the activation of SARS-CoV-2 S. Many betacoronaviruses including SARS-CoV-2 have polybasic residues at the S1/S2 site which is subjected to the cleavage by furin. The S1/S2 cleavage facilitates more assessable RBD to the receptor ACE2, and the binding triggers further conformational changes and exposure of the S2’ site to proteases such as type II transmembrane serine proteases (TTPRs) including TMPRSS2. In the presence of TMPRSS2 on the target cells, SARS-CoV-2 can utilize a direct entry route by fusion of the viral envelope to the cellular membrane. In the absence of TMPRSS2, SARS-CoV-2 enter target cells via endosomes where multiple cathepsins cleave the S for the successful entry. Additional host proteases involved in the cleavage of the S were discussed. This article also includes roles of 3C-like protease inhibitors which have inhibitory activity against cathepsin L in the entry of SARS-CoV-2, and discussed the dual roles of such inhibitors in virus replication.
Brucellosis remains one of the most common zoonoses spread worldwide, inducing enormous economic losses to the livestock industry and posing serious health threats to humans. Brucellosis re-emerged in China in the mid-1990s and reached a historically high level in 2015. The National Brucellosis Prevention and Control Plan (NBPCP) was initiated from 2016 to 2020. However, the present epidemiological status in livestock has not been elucidated, and whether Brucella variation occurred remains unclear. This study performed an extensive serological investigation in ruminant livestock from 2019 to 2021 in central Gansu Province, China. In total, 11,296 samples from 337 farms were collected to detect the specific antibodies of Brucella. The yearly average serological prevalence of Brucella at the flock level and individual level declined from 11.32% to 8.26% and 1.17% to 0.57%, respectively. The apparent individual-level seroprevalence of small and large ruminants was 0.89% and 0.52%, respectively. The brucellosis distribution has shifted from pastoral areas to agro-pastoral areas. Flock size and gender may be major risks of Brucella infection. Then, the B. melitensis TZ strain was isolated from female Tibetan sheep blood cell lysates. Phonotypical characterization demonstrated that it belongs to B. melitensis. biovar 3, and multilocus sequencing typing results indicated that it belongs to ST8. The whole genome and subsequent phylogenetic analysis demonstrated that the B. melitensis TZ strain is genetically more closely related to the B. melitensis QH61 strain. The B. melitensis TZ strain has similar growth characteristics to the B. melitensis 16 M strain. Overall, our study suggests that after strengthening control and prevention measures based on the NBPCP, there is a very low prevalence or absence of B. melitensis in the central Gansu Province of China, and the genotype of an epidemic strain of Brucella in Northwest China is relatively stable.
Coronaviruses are widespread in nature and can infect mammals and poultry, making them a public health concern. Globally, prevention and control of emerging and re-emerging animal coronaviruses is a great challenge. The mechanisms of virus-mediated immune responses have important implications for research on virus prevention and control. The antigenic epitope is a chemical group capable of stimulating the production of antibodies or sensitized lymphocytes, playing an important role in antiviral immune responses. Thus, it can shed light on the development of diagnostic methods and novel vaccines. Here, we have reviewed advances in animal coronavirus antigenic epitope research, aiming to provide a reference for the prevention and control of animal and human coronaviruses.
Rabies is an ancient disease. Two centuries since Pasteur, fundamental progress occurred in virology, vaccinology, and diagnostics—and an understanding of pathobiology and epizootiology of rabies in testament to One Health—before common terminological coinage. Prevention, control, selective elimination, and even the unthinkable—occasional treatment—of this zoonosis dawned by the twenty-first century. However, in contrast to smallpox and rinderpest, eradication is a wishful misnomer applied to rabies, particularly post-COVID-19 pandemic. Reasons are minion. Polyhostality encompasses bats and mesocarnivores, but other mammals represent a diverse spectrum of potential hosts. While rabies virus is the classical member of the genus, other species of lyssaviruses also cause the disease. Some reservoirs remain cryptic. Although global, this viral encephalitis is untreatable and often ignored. As with other neglected diseases, laboratory-based surveillance falls short of the notifiable ideal, especially in lower- and middle-income countries. Calculation of actual burden defaults to a flux within broad health economic models. Competing priorities, lack of defined, long-term international donors, and shrinking local champions challenge human prophylaxis and mass dog vaccination toward targets of 2030 for even canine rabies impacts. For prevention, all licensed vaccines are delivered to the individual, whether parenteral or oral–essentially ‘one and done’. Exploiting mammalian social behaviors, future ‘spreadable vaccines’ might increase the proportion of immunized hosts per unit effort. However, the release of replication-competent, genetically modified organisms selectively engineered to spread intentionally throughout a population raises significant biological, ethical, and regulatory issues in need of broader, transdisciplinary discourse. How this rather curious idea will evolve toward actual unconventional prevention, control, or elimination in the near term remains debatable. In the interim, more precise terminology and realistic expectations serve as the norm for diverse, collective constituents to maintain progress in the field.
Antibiotics are widely used to treat various diseases. However, growing evidence indicates that antibiotic therapy in human life increases the incidence of inflammatory bowel disease (IBD). Therefore, we need appropriate methods to reduce the incidence or symptoms of IBD. In this study, we used lincomycin hydrochloride to construct a gut microbial dysbiosis model in mice, and then, constructed an ulcerative colitis (UC) model. Meanwhile, we used Lactobacillus plantarum A3 from equine to treat UC in mice with gut microbial dysbiosis. The results showed that lincomycin hydrochloride had little effect on the small gut microbiota in mice, but had a more destructive effect on the large intestin. Lactobacillus plantarum A3 alleviated the symptoms of UC in mice, which was reflected in its significantly reduced spleen index and disease activity index (DAI) (p < 0.05), inhibited the shortening of colon and alleviated the invasion of inflammatory cells in the colon. Moreover, we found that it played a mitigatory role by inhibiting oxidative stress and regulating inflammatory cytokines in mice. At the same time, it restored the diversity and composition of the colonic microbiota and significantly increased the abundance of beneficial bacteria such as Blautia and Akkermansia (p < 0.05); Notably, it significantly increased the concentrations of arachidonoyl ethanolamide phosphate (AEA-P) and cortisone (p < 0.05) which have analgesic and anti-inflammatory effects. In conclusion, our study found that Lactobacillus plantarum A3 has the potential to regulate UC in mice with gut microbial dysbiosis.
The economic losses caused by uterine infections in ruminants have received increasing attention. Autophagy-related 7 (ATG7) has been demonstrated to be capable of regulating apoptosis, but surprisingly, ATG7 can both promote and inhibit apoptosis in different cellular contexts. However, the role of ATG7 in endometrial cell apoptosis during the postpartum period remains unclear. Herein, the location and expression of ATG7 was determined after mice were treated with lipopolysaccharide (LPS) in the uterus. The results showed that lipopolysaccharide (LPS) increased ATG7 expression in endometrial epithelial cells (EECs) but not endometrial stromal cells. The apoptosis of goat EECs was increased under LPS treatment, and LPS further led to an increase in bax expression and a decline in bcl-2 expression in goat EECs. Silencing of ATG7 inhibited goat EEC apoptosis with LPS treatment. The role of ATG7 in the regulation of goat EEC apoptosis was further confirmed by overexpression. ATG7 may serve as an essential regulatory factor in the process of endometrial epithelial cell apoptosis in ruminants under inflammatory injury. The findings help elucidate the pathogenesis of postpartum endometritis in ruminants.
Toxoplasma gondii (T. gondii) is an obligate intracellular parasite with a wide range of hosts, including humans and many warm-blooded animals. The parasite exists in two interconvertible forms, namely tachyzoites and bradyzoites in intermediate hosts that are responsible for acute and chronic infections respectively. Mature bradyzoites accumulate large amounts of amylopectin granules but their roles have not been fully characterized. In this study, the predicted key enzymes involved in amylopectin synthesis (UDP-sugar pyrophospharylase, USP) and degradation (alpha-glucan water dikinase, GWD) of ME49 strain were individually knocked out, and then bradyzoite-related phenotyping experiments in vitro and in vivo were performed to dissect their roles during parasite growth and development. Deletion of the usp or gwd gene in the type II strain ME49 reduced the replication rates of tachyzoites in vitro and parasite virulence in vivo, suggesting that amylopectin metabolism is important for optimal tachyzoite growth. Interestingly, the Δusp mutant grew slightly faster than the parental strain under stress conditions that induced bradyzoite transition, which was likely due to the decreased efficiency of bradyzoite formation of the Δusp mutant. Although the Δgwd mutant could convert to bradyzoite robustly in vitro, it was significantly impaired in establishing chronic infection in vivo. Both the Δusp and Δgwd mutants showed a dramatic reduction in the reactivation of chronic infection in an in vitro model. Together, these results suggest that USP and GWD, which are involved in amylopectin synthesis and degradation have important roles in tachyzoite growth, as well as in the formation and reactivation of bradyzoites in T. gondii.
The bacterium Avibacterium paragallinarum, previously known as Haemophilus paragallinarum, is responsible for causing infectious coryza (IC) in chickens and other avian species. In this case report, an outbreak of Avibacterium paragallinarum occurred in the Qinling area of China, resulting in clinical symptoms of facial swelling in several bird species, including Golden pheasant, Temminck's tragopan, and Peafowls, and three Golden pheasants died due to prolonged infection. Specific PCR results confirmed the presence of the pathogen in the infected birds. The report describes the clinical symptoms and pathological changes observed in the affected birds, as well as the isolation and identification of Avibacterium paragallinarum. Whole-genome sequencing and phylogenetic analysis were performed, and this is the first report of inter- and intra-species transmission of infectious coryza among wild birds in China.
A cross-sectional study was conducted to evaluate the risk factors associated with the Salmonella infection status of dairy herds in Henan and Hubei provinces, China. Herds were assigned a Salmonella status based on the isolation of Salmonella from fecal samples obtained from adult lactating dairy cows. Information on potential biosecurity risk factors was collected using a questionnaire template via a 15-min face-to-face interview with dairy cattle producers from May 2020 to March 2021. The questionnaire consisted of 23 questions across two broad categories of potential biosecurity risk factors: farm and biosecurity managemental factors. A total of 48 farms were surveyed. In all (100%, 48/48) of the surveyed farms, although sick animals were separated from healthy animals using fencing, there were no strict quarantine protocols for newly introduced animals. Mixed species rearing was practiced in 35.4% (17/48) of the farms. Feces were removed more than once a day in 45.8% (22/48) of the farms, whereas the remainder (54.2%, 26/48) only removed animal feces once a day. A total of 29.2% (14/48) of the farms were located < 2 km from other livestock farms. The use of personal protective equipment was consistently performed on all farms, with a majority of the workers on most farms (81.3%, 39/48) always disinfecting footwear before entering production areas. A significant association between Salmonella-positive status and the high frequency of the presence of wildlife (birds and rodents in sheds and feed storage places) was recorded (OR: 11.9, 95% CI: 1.7, 84.1, p = 0.013). Fortunately, no farms shared farm equipment with other farms. The study highlights the occurrence of wildlife as a risk factor for the presence of Salmonella in investigated dairy herds. There is a need to institute appropriate on-farm control measures for wild birds and rodents to control the potential spread of Salmonella in dairy production systems.
Variations in the pathogenicity of Newcastle disease virus (NDV), the agent causing Newcastle disease, are associated with variants of different virulence. A few studies have characterized the expression of microRNAs (miRNAs) in NDV-infected avian cells. Here, the expression of miRNAs in chicken embryo fibroblasts (CEFs) infected with Herts/33 and LaSota NDV strains (highly virulent and nonvirulent, respectively) was determined using RNA sequencing. miRNAs involved in NDV infection included 562 previously documented and 184 novel miRNAs. miRNA target genes involved transcription factors, cell apoptosis, ubiquitin-mediated proteolysis, and protein processing in the endoplasmic reticulum. Potential target genes associated with autophagy were verified by qRT-PCR. No studies have documented the miRNA profiles of CEFs infected with NDVs variants. This study adds to our knowledge of the cellular miRNAs involved in NDV infection and the complex molecular mechanisms mediating virus-host interactions. The results of this study will aid the development of strategies against the chicken virus.
Broiler ascites syndrome (AS) is one of the main diseases threatening the health of broilers. It is well documented that myocardial hypertrophy and failure is one of the key mechanisms of broiler ascites syndrome. Therefore, prevention of cardiac hypertrophy and failure would be one goal to reduce broiler ascites syndrome incidence. Myocardial hypertrophy and failure are closely related to endoplasmic reticulum stress (ERS) in cardiac myocytes, and the endoplasmic reticulum stress signaling system (ATF6-DR5) is one of the important pathways of myocardial apoptosis. Excessive hypertrophy will affect the heart muscle’s normal contraction and diastole function, and the heart will turn from compensated to decompensate thus causing myocardial injury. Myocardial apoptosis is a core component of the pathological changes of this myocardial injury. Nano-selenium is a kind of red elemental selenium nanoparticle. Due to its excellent physical, chemical and biological properties, it has attracted extensive academic attention in recent years. It has been proven to have excellent antioxidant, antibacterial, antitumor, antihypertrophic, and antiapoptotic abilities. Herein, nano-selenium (1 µmol/L) can inhibit hydrogen peroxide (H2O2)-induced oxidative stress in broiler primary cardiomyocytes, and at the same time reduce cardiomyocyte apoptosis. In vivo, nano-selenium can reduce broiler myocardial injury-related enzyme indicators (AST, CK and LDH), and alleviate myocardial injury. It can also activate the antioxidant enzyme system (SOD, GSH-Px and CAT) and reduce MDA, and make the recovery of T-AOC ability in the organization. Meanwhile, nano-selenium can down-regulate the genes and proteins expression of ATF-6, GRP-78, CHOP and caspase 12 in the ERS-related signaling pathway, and inhibit that of downstream-related caspase 3, Bax and caspase 9, and increase that of the downstream anti-apoptotic Bcl-2, thereby maintaining the homeostasis of the endoplasmic reticulum and alleviating cardiomyocyte apoptosis. It can be seen that nano-selenium can protect the damaged myocardium in the broiler ascites caused by high-salt drinking by regulating the ATF6-DR5 signaling pathway. This study was performed in chickens and cardiomyocyte cells and attempted to demonstrate that selenium nanoparticles can protect the damaged myocardium in broiler ascites. This paper provides a new idea for preventing and treating broiler ascites syndrome.
The regulation and maintenance of bone metabolic homeostasis are crucial for animal skeletal health. It has been established that structural alterations in the gut microbiota and ecological dysbiosis are closely associated with bone metabolic homeostasis. The gut microbiota and its metabolites, especially short-chain fatty acids (SCFAs), affect almost all organs, including the bone. In this process, SCFAs positively affect bone healing by acting directly on cells involved in bone repair after or by shaping appropriate anti-inflammatory and immunomodulatory responses. Additionally, SCFAs have the potential to maintain bone health in livestock and poultry because of their various biological functions in regulating bone metabolism, including immune function, calcium absorption, osteogenesis and osteolysis. This review primarily focuses on the role of SCFAs in the regulation of bone metabolism by gut microbiota and provides insight into studies related to bone health in livestock and poultry.
Transmissible spongiform encephalopathies (TSEs) are a group of progressive and ultimately fatal neurologic diseases of man and animals, all resulting from the propagated misfolding of the host’s normal cellular prion protein. These diseases can be spontaneous, heritable, anthropogenic/iatrogenic, or in some cases horizontally transmissible, and include such notable TSEs as bovine spongiform encephalopathy (BSE) of cattle and chronic wasting disease (CWD) of cervids. Although they are both unequivocally protein misfolding disorders, they differ markedly in their pathogenesis, transmissibility, and zoonotic potential. While the BSE epidemic has largely abated over the past three decades following global feed bans on ruminant meat and bone meal, CWD, which is readily transmitted through various forms of excreta, has rapidly expanded from its original endemic zone to encompass much of North America, along with recently identified foci in Scandinavia. Most importantly, although the classical form of BSE has proven transmissible to humans consuming contaminated beef or beef products, so far there have been no conclusive reports on the zoonotic transmission of CWD to humans. The underlying basis for these differences – whether host or agent directed – are not well understood, though may be due to inherent differences in the three-dimensional structure of the misfolded BSE or CWD prion proteins or the expression levels and tissue distribution of respective cellular prion proteins. With the uncontrolled geographic spread of CWD, it is imperative that we improve our understanding of the factors governing prion disease pathogenesis, transmission, and zoonotic potential.
Spillover of trypanosomiasis parasites from wildlife to domestic livestock and humans remains a major challenge world over. With the disease targeted for elimination by 2030, assessing the impact of control strategies in communities where there are human-cattle-wildlife interactions is therefore essential. A compartmental framework incorporating tsetse flies, humans, cattle, wildlife and various disease control strategies is developed and analyzed. The reproduction is derived and its sensitivity to different model parameters is investigated. Meanwhile, the optimal control theory is used to identify a combination of control strategies capable of minimizing the infected human and cattle population over time at minimal costs of implementation. The results indicates that tsetse fly mortality rate is strongly and negatively correlated to the reproduction number. It is also established that tsetse fly feeding rate in strongly and positively correlated to the reproduction number. Simulation results indicates that time dependent control strategies can significantly reduce the infections. Overall, the study shows that screening and treatment of humans may not lead to disease elimination. Combining this strategy with other strategies such as screening and treatment of cattle and vector control strategies will result in maximum reduction of tsetse fly population and disease elimination.
Canine atopic dermatitis (CAD) is a prevalent genetically susceptible inflammatory and pruritic allergic skin condition affecting not only the health of dogs but also the quality of life of their owners. Interleukin-31 (IL-31) and interleukin-31 receptor alpha (IL-31RA) are essential for the development of pruritus in primates and mice. Hence, it is expected that inhibiting IL-31RA will be an effective approach to alleviate pruritus. The purpose of the study was to produce anti-canine IL-31RA polyclonal antibodies (anti-IL-31RA pAbs) and evaluate their efficacy in inhibiting house dust mite (HDM)-evoked pruritic responses. Dogs were immunized with antigens formed by IL-31RA recombinant short peptides coupled to BSA to produce anti-IL-31RA pAbs. The CAD model was developed by using HDM allergen stimulation, and the effects of IL-31RA pAbs on the reduction of pruritus in CAD model dogs were examined. The Canine Atopic Dermatitis Extent and Severity Index (CADESI)-4 and pruritus Visual Analog Scale (pVAS) were utilized to evaluate pruritic responses, and skin tissue samples were collected from the inguinal area for pathological assessment of skin inflammatory cell infiltration. The results showed that anti-IL-31RA pAbs with high titers (1:128,000) and specificity were effectively produced. In the CAD model group, the severity of skin damage, pruritus score, inflammatory cell infiltration and level of inflammatory factors were considerably elevated. Anti-IL-31RA pAbs relieved pruritic behavior and dermatitis in dogs compared to placebo-treated dogs. In conclusion, anti-IL-31RA pAbs effectively suppressed CAD in vivo and are anticipated to be an effective novel treatment for pruritic skin disorders such as CAD.
Bullfrogs (Rana catesbeiana) are amphibians with high economic value, but in recent years, bullfrog farming has encountered serious threats of bacterial diseases, and the “bullfrog economy” is facing a continuous decline. In this study, the dominant strain was isolated from diseased bullfrogs in a bullfrog farm in Nanning, Guangxi, and based on its morphological, physiological, and biochemical characteristics and analysis of 16S rRNA gene sequences, the strain was identified as a non-O1/O139 group Vibrio cholerae and named TC1. Three virulence factors were identified in this strain, including hemolysin, outer membrane protein, and toxin-coregulated pili. Drug susceptibility testing showed that the strain resisted gentamicin, florfenicol, nitrofural, oxytetracycline, neomycin, penicillin, amoxicillin, doxycycline, and sulfamonomethoxine. The results of artificial infection experiments showed that TC1 caused serious pathologies such as abdominal swelling and anal prolapse in bullfrogs, especially severe intestinal bleeding. Histopathological observations revealed that the bullfrog intestine exhibited obvious pathological lesions. These results provide an essential epidemiological basis for controlling V. cholerae infections in aquatic animals and demonstrate the promise of bullfrogs as an amphibian model for studying the pathogenesis of V. cholerae.
Toxoplasma gondii is thought to infect all nucleated cells in warm-blooded animals, including poultry, mammals, and humans. However, it is unclear whether T. gondii can infect chicken erythrocytes due to the nucleated nature of these cells. Due to the special role of chicken erythrocytes in innate immunity, we investigated the cell‒cell interaction between T. gondii and erythrocytes to elucidate the role of chicken erythrocytes in T. gondii infection. Cellular apoptosis was analyzed by transwell assay and flow cytometry. An immunofluorescence method was used to examine the reorganization of vimentin during T. gondii infection in both Vero cells and chicken erythrocytes. The reorganization of actin was evaluated to further examine the invasion capacity of tachyzoites on chicken erythrocytes during infection. We discovered that T. gondii can adhere to but not invade chicken erythrocytes and eventually cause apoptosis in chicken erythrocytes. When tachyzoites were cocultured with chicken erythrocytes in vitro, the transcriptional levels of T. gondii MIC3, ROP16, and ROP18 were significantly decreased. In addition, the rearrangement of host cell vimentin, a type III cytoskeleton protein regulated by T. gondii infection, was not observed. Similarly, the parasite-induced ring-shaped actin structure was not formed in the host-parasite junction. T. gondii (RH strain) tachyzoites preferentially invaded Vero cells and replicated in chicken blood monocytes, but they were not found in chicken erythrocytes. These findings showed that although T. gondii could attach to the surface of chicken erythrocytes, but couldn’t invade successfully. Interestingly, we found that the T. gondii secretome, lysates, and intact tachyzoites could cause apoptosis of chicken erythrocytes, which suggested a complex mechanism involved in the apoptosis of chicken erythrocytes induced by T. gondii. This study elucidated that T. gondii could not infect nucleated chicken erythrocytes and enriched our understanding of the transmission mechanism of T. gondii among avian species.
In the global progress of bone tumor research, established stable and long-lasting transgenic chondrosarcoma (CSA) cell lines are rare, mainly of murine and human origin, while the establishment of canine CSA cell lines has yet to be reported. This study established a canine CSA cell line to facilitate the basic clinical study of canine CSA. Fifty five cases of canine osteolytic disease were collected, and more than 10 bone tumor samples from dogs with typical clinical signs were used for primary cell culture. A cell line with stable passaging for more than 100 generations and mouse tumorigenic ability was successfully cultured. According to the clinical characteristics of the dog and the histopathological results of the primary tumor, CSA was diagnosed, and the CSA cell line was designated Mango. Immunohistochemical (IHC) results showed that the immunoreactivity of bone gamma-carboxyglutamate protein (BGLAP), secreted protein acidic and rich in cysteine (SPARC), alkaline phosphatase (ALPL), vimentin (VIM) and S100 were positive. However, the immunoreactivity of pan-cytokeratin (PCK), chromogranin A (CGA), and platelet endothelial cell adhesion molecule-1 (CD31) was negative. Immunofluorescence (IF) results showed that the protein expressions in the Mango cell line were consistent with the IHC identification of the primary tumor. The Mango cell line’s doubling time was 43.92 h, and the cell formation rate exceeded 20%. There were abnormal chromosome numbers, hetero staining with toluidine blue, and certain calcification abilities. It could be passaged stably and continuously without changing the cell morphology and characteristics. In vivo, the cells were successfully injected into the nude mice model with a tumorigenic rate of 100%. The immunophenotype of the xenograft tumor was consistent with that of the primary tumor. Therefore, we effectively established a canine CSA cell line. As a promising cell material, this cell line can be used to construct a tumor-bearing model conducive to the subsequent basic research of canine CSA. Moreover, because of its similarity to human CSA, the animal model of CSA is also indispensable for investigating human CSA.
Pulmonary macrophages, such as tissue-resident alveolar and interstitial macrophages and recruited monocyte-derived macrophages, are the major macrophages present in the lungs during homeostasis and diseased conditions. While tissue-resident macrophages act as sentinels of the alveolar space and play an important role in maintaining homeostasis and immune regulation, recruited macrophages accumulate in the respiratory tract after acute viral infections. Despite sharing similar anatomical niches, these macrophages are distinct in terms of their origins, surface marker expression, and transcriptional profiles, which impart macrophages with distinguished characteristics in physiological and pathophysiological conditions. In this review, we summarize the current view on these macrophage populations, their shared functions, and what makes them distinct from each other in the context of homeostasis and respiratory viral infections.
Retroviruses have been proven to cause infections and diseases in a series of mammalian hosts but not in dogs. Then, this letter discussed the dog susceptibility to retrovirus infection, encompassing arguments to understand why dogs may have not been infected by retroviruses thus far. The potential resistance of retrovirus in dogs enables this provocative short communication to discuss this question, looking at some evolutive aspects. The lineage of canids has shown, throughout its evolutionary history, a smaller accumulation of retroviruses in canid genomes, classified as endogenous retroviruses. In this context, the genomes of canids seem to offer obstacles, which have been evolutionarily conserved, in the face of retroviral infection.
Hematology, plasma biochemistry, body composition, and bone mineral density (BMD) were analyzed for 30 captive Ceratophrys cranwelli (C. cranwelli) to establish the reference intervals. There was no significant difference between males and females in all blood routine tests. Blood biochemistry included 17 analytes, and only total bile acid (TBA), calcium (CA), and phosphorus (PHOS) showed significant differences. Male TBA levels were higher than females, while female CA and PHOS levels were higher than males. The body composition and BMD of males and females were similar, except for bone area, which showed a significant gender difference, with females having higher values than males. The data obtained in this study can help with the medical management of diseased individuals and serve as a reference for health assessments of future populations.
Salmonella enterica has a wide diversity, with numerous serovars belonging to six different subspecies with dynamic animal-host tropism. The FimH protein is the adhesin mediating binding to various cells, and slight amino acid discrepancy significantly affects the adherence capacities. To date, the general function of FimH variability across different subspecies of Salmonella enterica has not been addressed. To investigate the biological functions of FimH among the six Salmonella enterica subspecies, the present study performed several assays to determine biofilm formation, Caenorhabditis elegans killing, and intestinal porcine enterocyte cell IPEC-J2 adhesion by using various FimH allele mutants. In general, allelic mutations in both the lectin and pilin domains of FimH could cause changes in binding affinity, such as the N79S mutation. We also observed that the N79S variation in Salmonella Dublin increased the adhesive ability of IPEC-J2 cells. Moreover, a new amino acid substitution, T260M, within the pilin domain in one subspecies IIIb strain beneficial to binding to cells was highlighted in this study, even though the biofilm-forming and Caenorhabditis elegans-killing abilities exhibited no significant differences in variants. Combined with point mutations being a natural tendency due to positive selection in harsh environments, we speculate that allelic variation T260M probably contributes to pathoadaptive evolution in Salmonella enterica subspecies IIIb.
Management strategies and the use of advanced technologies are equally important for determining the sample number and sampling frame for successful field sampling for animal disease prevalence studies. The quality of the biological samples collected in the field has a direct bearing on the integrity of the data generated, prevalence estimates and subsequent policy decisions on disease control. Hence, compromising the quality of biological samples collected in the field could potentially undermine the priority setting principles in disease control strategies. Biological samples collected from domestic animals in the field are precious materials and require meticulous planning for sample collection, sample storage in the field, transportation, and storage in the laboratory. Poorly managed field sampling has a significant detrimental impact on the sample quality and quantity and directly affects the accuracy of disease prevalence data. A bad choice of sampling tools, containers, storage and transport all have a negative impact on the integrity of the sample and consequently have an impact on the outcome. Over the last two years, as part of our one health animal sampling work in India, we have observed challenges and opportunities in the field sampling of animals for disease prevalence studies. This paper aims to provide information on management practices and technologies for efficient biological sample collection from the field and ensure that good quality samples are available for testing.
The rapid expansion of large-scale pig farming has brought about a surge in viral diseases with high morbidity rates and diverse manifestations. This widespread occurrence of multiple viral diseases in pig farms has inflicted severe economic losses on the global swine industry. Consequently, there is an urgent need for eco-friendly and efficient antiviral drugs that can effectively combat viruses and prevent diseases such as PEDV, PRRSV, PRV, and other viral infections. To this end, we conducted a study on the antiviral activity and cytotoxicity of eleven different Chinese herbal extracts (CHE) against PRV. In vitro testing of several extracts, namely, Echinacea, Ilex purpurea Hassk, Ganoderma lucidum Kars, Taraxacum mongolicum, and Ilex rotunda Thunb, exhibited remarkable inhibition of PRV infection without causing any cytotoxic effects. Specifically, their antiviral selectivity indexes were significantly higher, with values ranging from 6- to 144-fold. The antiviral efficacy of five CHEs was evaluated against other RNA viruses, including PRRSV and PEDV. The extracts showed substantial inhibition of PEDV and PRRSV proliferation. Echinacea and Ilex purpurea Hassk extracts exhibited the highest virus inhibitory effects. To understand the antiviral mechanisms underlying their potent activity, a time-of-addition experiment was conducted. The results indicated that these extracts effectively targeted the early infection and postinfection stages of PRV, PEDV, and PRRSV. The study found that the Chinese herbal extracts, Echinacea and Ilex purpurea Hassk, had both direct and indirect effects on virus particles and cellular targets, demonstrating broad-spectrum antiviral activity against multiple clinical strains of PRV and PEDV. These findings provide a strong foundation for the development of herbal medicines to prevent and treat infections caused by PRV, PEDV and PRRSV in the swine industry. The identified extracts show great promise for the formulation of effective and environmentally friendly antiviral interventions.
The clinical application of solid lipid particles (SLPs) is hampered due to the need for advanced nano/micro-suspension production technology. This research aims to establish a pilot-scale production line employing high-speed shears as emulsification equipment. The primary purpose is to manufacture nano/micro-suspensions using solid lipid particles (SLPs). The study also exhaustively introduces and analyzes the regulatory schemes for process parameters and formulations at various stages of production. The process and formulation endured optimization through orthogonal or single-factor tests at various production steps: laboratory research, small-scale trial production, and pilot production. Quality standards for the product were determined, and key parameters were obtained at each stage. The laboratory research demonstrated that the optimal SLPs comprised 15 mL 3% polyvinyl alcohol (PVA) per 1.0 g tilmicosin and 2.5 g carnauba wax (WAX). During small-scale production, modifications were made to the volume of the aqueous phase, emulsifier concentration, and emulsification strength, setting them to 16 mL, 5%, and 2200 r/min, respectively. In the pilot production stage, the shear time was considered optimal at eight min. The impurity, content, polydispersion coefficient (PDI), and size of the pilot product were < 3%, 5%, 0.385 and 2.64 μm, respectively. Among the several parameters studied, heating temperature, drug-lipid ratio, and emulsifier concentration were identified as the main factors affecting product quality, and they were regulated at 100℃, 1:3, and 5%, respectively. A novel hot melt emulsification shear method aided the development of a new solid lipid-based suspension from its preliminary stages in the laboratory to pilot production. This innovation is expected to enhance solid lipid-based suspensions' industrial evolution extensively.
Chronic kidney disease is commonly diagnosed in dogs, and clinical signs may be aggravated when infected agents are involved. In this case report, 33 dogs with chronic kidney disease were clinically evaluated and serologically tested for Leptospira spp., Ehrlichia canis, and Anaplasma phagocytophilum. The seroprevalence for Leptospira spp. was 39.4%. The most frequent serovars found were Pyrogenes, Canicola, Bratislava and Australis, with serological titers between 1:100 to 1:800. Clinical signs included fever, depression, decreased body condition, vomiting and hematuria. Significant laboratory findings were anemia, leukocytosis, thrombocytopenia, increased liver enzymes, urea and creatinine, hyperbilirubinemia and hyperphosphatemia. All leptospira seronegative dogs were positive for one or both monitored homoparasites (i.e., E. canis and A. phagocytophilum); only three leptospira seropositive dogs were positive for one or both hemoparasites. Findings also suggest that endemic hemoparasites of dogs should be monitored in dogs with a kidney condition for a better clinical picture of the patients and therapeutic approach.
H9N2 avian influenza viruses (AIVs) are widely distributed, causing continuous outbreaks in poultry and sporadic infections in humans. Vaccination is the primary method used to prevent and control H9N2 AIV infection. However, the ongoing evolution and mutation of AIVs often result in limited protection effects from vaccines. Therapeutic monoclonal antibodies (mAbs) targeting influenza viruses offer a promising alternative. In this study, we immunized mice with inactivated H9N2-W1 virus, and we screened and acquired five mAbs, namely 4D12, F4, 5C8, 2G8 and A11. We showed that all five mAbs specifically targeted the HA protein of various H9N2 AIV strains. In vitro neutralization tests demonstrated that all five mAbs exhibited neutralization activity against H9N2 AIVs, with mAb F4 displaying the most potent neutralization effect. The F4 mAb exhibited dose-dependent preventive and therapeutic effects against lethal H9N2-115 infection, and the administration of F4 at a dose of 3 μg/g provided complete protection in vivo. Our study presents an alternative approach for preventing and controlling H9N2 AIV infection. Furthermore, the identified F4 mAb holds promise as a solution to potential pandemics in humans caused by H9N2 AIVs.
Foot-and-mouth disease (FMD) is a highly contagious viral disease affecting cloven-hoofed livestock. It is caused by the FMD virus (FMDV), which has seven distinct serotypes (O, A, C, SAT I, SAT II, SAT III, and Asia 1). In Nepal, FMD is a prevalent and economically important livestock disease, with hundreds of outbreaks yearly across different regions. However, there is limited understanding of the recent epidemiological trends of FMD in the past few years in Nepal. This study aims to analyze the spatial and temporal distribution of FMD in Nepal from 2019 to 2021. The FMD and TADs Investigation Laboratory, under the Government of Nepal, conducts annual risk-based surveillance of FMD in the country. The nonstructural protein (NSP) serosurveillance and serotyping (for outbreak confirmation) data from this laboratory were used for the study. The samples were collected either by the laboratory staff or were sent to the laboratory. Data analysis and mapping were performed using Epi info version 7.2.5.0 and QGIS version 3.22.5, respectively. Our findings revealed that 37.65% of samples (n = 417) tested positive for serotyping. The highest number of positive cases occurred in March–April, followed by December. Geographically, the Terai region had the most positive cases, followed by hills and mountains. The positivity rate for serotyping did not significantly vary by animal species (p > 0.05). Serotype O was the dominant serotype in all years, accounting for 98% of cases, while serotype A was found in only 2% of serotype-positive samples. In NSP serosurveillance, out of 3216 samples tested, 15.07% (474/3146; 95% CI, 13.86–16.36) tested positive. NSP seropositivity varied significantly by year (p < 0.001) but not by eco-zone (p > 0.05). In conclusion, FMD remains endemic in Nepal, with a consistent epidemiological pattern except that the Asia 1 serotype was not detected in the past years. We recommend expanding FMD surveillance activities to high-risk areas and collecting data on potential risk factors driving FMD infection in the country. This will enable the implementation of suitable control measures.
Despite the initial successes of the Bacillus Calmette-Guerin (BCG) vaccine in children, its efficacy against tuberculosis is highly variable. There is a lack of understanding about how mental conditions influence BCG vaccination. Here, we used the chronic social defeat stress (CSDS) model to explore the effects of depression on BCG vaccination efficacy. We observed higher lung and spleen bacterial loads and a lower organ index in depressed compared to BCG mice. Meanwhile, a relatively lower T cell protective efficacy was observed in both compared to control and BCG mice via a mycobacterium growth inhibition assay (MGIA). Cytokine expression of IL-12p40, IL-1β, IL-17, TNF-α and IFN-γ was reduced, whereas the expression of IL-10 and IL-5 was increased in the spleen of both compared to BCG mice. Moreover, the proportions of CD4+IFN-γ+, CD8+IFN-γ+ T lymphocytes and CD4+ effector/central memory T cells were reduced in the splenocytes of the depressed BCG mice. Depression promotes CD4+ regulatory T cells (Treg) and myeloid-derived suppressor cell (MDSC) generation in depressed mice, contributing to the reduced pro-inflammatory immune response upon BCG vaccination. This study provides insight into the decreased protective immunity by BCG vaccination attributable to depression in mice.