A methodology designed to eliminate mitotic inhibitor action and involving use of pretested fetal calf serum and careful pH and temperature control has been described by which cells from normal human and animal tissue can be maintained in active growth for long periods in vitro without development of aneuploidy. By means of this procedure, it is possible reliably to establish cell cultures from minute skin biopsies which can be taken from any individual. Clones of mammalian cells with chromosomal markers have been isolated by this means from x-irradiated non-irradiated cell cultures. Application of these techniques to chromosome delineation in large numbers of human subjects; determination of chromosomal sex in patients; spontaneuos and induced genetic changes in somatic mammalian cells in vivo and in vitro; comparison of metabolic differences between normal and cancerous cells and other problems have been indicated.
Wildlife rescue centres face considerable challenges in promoting animal welfare and enhancing the care and housing conditions of animals under professional supervision. These challenges are further compounded by the diversity of species admitted, each with distinct specific needs. In Colombia and other Latin American countries, primates are among the most frequently rescued and behaviourally complex mammalian taxa, requiring particular attention. In response, this study aimed to assess the content validity of proposed animal welfare indicators for <i>Cebus albifrons</i> through a Delphi consultation process and to develop two species-specific assessment protocols: a daily-use tool for keepers and a comprehensive protocol for professional audits. A panel of 23 experts in primate care and rehabilitation participated in two consultation rounds to evaluate and prioritise the indicators based on their content validity, perceived reliability, and practicality. Indicators were classified as either animal-based (direct measures) or resource- and management-based (indirect measures). After each round, experts received summarised feedback to refine their responses and facilitate consensus building. Of the 39 initially proposed indicators, 28 were validated for inclusion in the extended protocol and 10 selected for the daily-use checklist. Among these, 20 indicators in the extended protocol and 6 in the daily protocol were resource- or management-based—such as adequate food provision, physical enrichment, and habitat dimensions—highlighting their practical applicability and relevance in identifying welfare issues and risk factors. Although these indirect indicators were more numerous, the top-ranked indicators in both protocols were animal-based, including signs of pain, affiliative behaviours, and abnormal repetitive behaviours. These are essential for accurately reflecting the animals’ welfare state and are therefore critical components of welfare assessment in captive non-human primates. This study demonstrates that welfare assessment tools can be effectively tailored to the specific needs of wildlife rescue centres, providing a robust foundation for enhancing welfare practices. These protocols not only offer practical approaches for assessing welfare but also underscore the importance of embedding animal welfare as a priority alongside conservation efforts. Future research should aim to refine these tools further, assess their implementation, and evaluate inter- and intra-observer reliability to ensure consistency across different settings.
Cesar Guillermo Meneses-Nava, Alessandra Ubiali, Greta Dossi
et al.
IntroductionFlow cytometry (FC) is widely used in humans and dogs to diagnose and characterize hematopoietic neoplasms. Conversely, its use in feline patients is still limited, leading to a lack of standardized protocols and subjective data interpretation.MethodsHerein, we describe FC features of circulating lymphoid subsets in a total of 20 cats: 9 healthy cats, 6 diseased cats without hematopoietic neoplasm, and 5 cats with probable chronic lymphocytic leukemia (CLL), using a panel of 10 antibodies and a multicolor approach, in terms of both cell size (nFSC) and degree of antigen expression (MFI).ResultsThree main subsets were identified in healthy cats and diseased cats without hematopoietic neoplasm (namely, CD5 + CD45R-, CD21 + CD45R + and CD5 + CD45R+). CD4 + CD8- cells outnumbered CD4- CD8 + cells. Low percentages of CD4 + CD8 + and CD134 + cells were also present. MHCII had higher fluorescence intensity in B- than in T-cells. CD9 was not expressed on leukocytes surface, but on small events possibly referable to platelet clumps. In diseased cats without hematopoietic neoplasm, each T-cell subset was larger in size than in healthy cats. Finally, in cats with probable CLL the leading phenotype was CD5 + CD45R-CD4 + CD8-CD134 + MHCII+ and cell size overlapped with the one of the other diseased cats.DiscussionOur results are expected to lay the ground for a more standardized approach to feline samples for FC, and a more objective data interpretation, ultimately leading to improved diagnostic accuracy. Further studies are needed to assess the biological, diagnostic and prognostic value of specific FC patterns in feline medicine.
ABSTRACT Bunyaviruses, a subset of segmented negative-sense RNA viruses, include pathogenic species capable of zoonotic transmission to humans via arthropod vectors and rodent hosts. Pathogenic bunyavirus infections can cause severe hemorrhagic fevers and other life-threatening diseases, posing threats to human health and social stability; however, therapeutic strategies for treating bunyavirus infections remain limited. Here, we report that VV251 hydrochloride salt (VV251), an optimized oral prodrug derivative of 4′-fluorouridine (EIDD-2794), exhibits potent efficacy against severe fever with thrombocytopenia syndrome virus (SFTSV) and lymphocytic choriomeningitis virus (LCMV) both in vitro and in vivo. In various cell lines, VV251 inhibits SFTSV and LCMV with EC50 values in the nanomolar to micromolar range. In lethal rodent models, once-daily oral administration of VV251 at low doses (10 mg/kg for SFTSV; 1 mg/kg for LCMV) achieves complete protection (100% survival), matching the efficacy of T-705 at 300 mg/kg. Additional pharmacokinetic analysis indicates that VV251 has favorable absorption and exposure profiles in both Sprague-Dawley rat and cynomolgus monkey models. This study evaluates the antiviral profile of VV251 and supports its further development as a promising therapeutic candidate.IMPORTANCEBunyaviruses encompass numerous highly pathogenic agents that pose significant threats to human health, including the causative agents of Crimean-Congo hemorrhagic fever, Lassa fever, and Rift Valley fever. The World Health Organization has identified Lassa fever as a priority pathogen requiring urgent research and development efforts in emergency contexts, underscoring the critical need for effective oral antiviral therapies to enhance pandemic preparedness. Here, we report that VV251 hydrochloride salt (VV251), an optimized oral prodrug derivative of 4′-fluorouridine (4′-FlU, EIDD-2794), shows significant efficacy against severe fever with thrombocytopenia syndrome virus and lymphocytic choriomeningitis virus infections, with inhibitory activity in cell culture and protective effects in lethal animal models. Building on the established broad-spectrum antiviral activity of 4′-FlU against multiple high-consequence pathogens (including severe acute respiratory syndrome coronavirus 2, respiratory syncytial virus, Lassa virus, and Junin virus), VV251 emerges as a promising next-generation oral antiviral candidate, offering an orally available therapeutic option to combat these formidable pathogens.
<p>The environment in which animals are kept must provide suitable conditions for their species. This includes ensuring that animals are healthy, well-fed, safe, able to exhibit species-specific behaviors, not experiencing fear or pain, and not under chronic or acute stress. Poultry welfare is achieved when birds are raised in environments that meet their physiological and ethological needs. Fear can significantly impact animal welfare. Chickens have been significantly altered by human artificial selection. Despite this, they exhibit reactivity towards humans and tend to avoid them. Poultry animals reared in environmentally controlled poultry houses and bred for superior productivity are more sensitive to fear factors and have lost their adaptability to a great extent. This study aimed to determine the effect of personnel clothing color on stress and fear in chickens in layer hen coops. The experiment involved 32-week-old laying hens of three different genotypes. A worker in the henhouse wore six respective different colors of workwear (dark blue, green, red, yellow, black, and white), and sound measurements were taken during this time. The results showed that the color of the worker's clothing influenced the sound intensity of the chickens (<span class="inline-formula"><i>P</i><i><</i>0.05</span>). White clothing elicited the least reaction, whereas black and dark blue elicited the most. The other three colors showed similar reactions. In conclusion, workers in layer hen coops wearing dark clothing, such as dark blue and black, can induce stress and noise in the animals. Additionally, chickens showed similar reactions to green, red, and yellow colors, with white being the color around which they felt the most secure.</p>
Abstract Chicken production in Tanzania provides opportunity to local communities in terms of employment, increased income, food security, and manure for cropping. However, diseases like fowl typhoid remain a challenge to livestock keepers. This study was aimed at understanding the attitude and awareness of Poultry keepers about the prevalence of fowl typhoid in chickens kept in Dodoma. A cross-sectional survey using semi-structured interviews was employed to understand farmers’ perception of the prevalence of fowl typhoid and associated risk factors amongst poultry farmers in three wards in Dodoma, namely, Nkuhungu, Msalato, and Mnadani. The overall prevalence of fowl typhoid among farmers was 22.30%, with significant differences being noticed in the first quarter (January–March) and the third quarter (July–September) (P < 0.05). Factors such as age and sex, flock size, and management practices influence the prevalence of fowl typhoid significantly (P < 0.05). Furthermore, the farmers had challenges accessing the veterinary services due to their unawareness, the availability of the service, and their distance from the service. The control strategies for fowl typhoid should consider the influencing factors while improving the accessibility and availability of veterinary services to farmers.
ABSTRACT: Circular RNAs (circRNAs) are a class of endogenous non-coding RNAs that have been implicated in mediating granulosa cell (GC) proliferation and apoptosis. CircRAB11A was found to have a significantly higher expression in normal follicles compared to atrophic follicles. In this study, we determined that the knockdown of circRAB11A resulted in the inhibition of proliferation and promotion of apoptosis in GCs of chicken. Moreover, circRAB11A was found to act as a sponge for miR-24-5p, both member RAS oncogene family (RAB11A) and epidermal growth factor receptor (EGFR) were revealed to be targets of miR-24-5p through a dual-luciferase reporter assay. RAB11A or EGFR promoted proliferation and suppressed apoptosis in GCs through the phosphatidylinositol-kinase (PI3K)/AKT or extracellular signal-regulated kinase (ERK)1/2 pathway. These findings suggest that circRAB11A may function as a competing endogenous RNA (ceRNA) by targeting the miR-24-5p/RAB11A and miR-24-5p/EGFR axes and activating the ERK1/2 and PI3K/AKT pathways, offering a potential avenue for exploring the mechanism of follicle development.
ABSTRACT: The current study was conducted to evaluate the effects of different administration routes of bacterial lipopolysaccharide (LPS) on intestinal mucosal morphological, immunological, and microbial barrier functions in goslings. First, we compared intestinal villi morphology of goslings under intraperitoneal or oral LPS treatment through hematoxylin and eosin staining. Then, we determined the signatures of the microbiome in the ileum mucosa of goslings subjected to oral LPS treatment at 0, 2, 4, and 8 mg/kg BW by 16S sequencing, and analyzed the changes in intestinal barrier functions and permeability, levels of LPS in the ileum mucosa, plasma, and liver tissue, and the induced inflammatory response of Toll-like receptor 4 (TLR4). As a result, intraperitoneal LPS injection resulted in a thicker intestinal wall in the ileum within a short time, whereas villus height was less affected; in contrast, oral LPS treatment exerted a stronger influence on villus height but not on intestinal wall thickness. We also found that oral LPS treatment affected the structure of the intestinal microbiome, reflected by changes in the clustering of intestinal microbiota. The average abundance of Muribaculaceae showed an increasing trend with increasing LPS levels, and that of the genus Bacteroides decreased, compared with the control group. In addition, oral LPS treatment with 8 mg/kg BW affected the intestinal epithelial morphology, damage the mucosal immune barrier, downregulated the expression of tight junction proteins, increased circulating D-lactate levels, and stimulated the secretion of various inflammatory mediators and activation of the TLR4/MyD88/NFκB pathway. This study presented the injuries of intestinal mucosal barrier function induced by LPS challenges in goslings and provided a scientific model for searching the novel strategies to attenuate the immunological stress and gut injury caused by LPS.
Arthropod-borne infections, known as vector-borne diseases, are a significant threat to both humans and animals. These diseases are transmitted to humans and animals through the bites of infected arthropods. In the last half century, there have been a number of unexpected viral outbreaks in Middle Eastern countries. Recently, Iraq has witnessed an outbreak of the Crimean-Congo Hemorrhagic Fever virus with high morbidity and mortality rates in humans. However, very little is known about the prevalence and distribution of CCHFV in Iraq, and therefore, it is impossible to quantify the risk of infection. CCHFV is transmitted to humans through the bite of infected ticks. However, transmission can also occur through contact with the blood or tissues of infected animals during slaughter or through contact with contaminated objects or surfaces. Accidental crushing of infected ticks can also release the virus and lead to transmission. In addition, healthcare workers and family members caring for infected individuals are at risk of infection through exposure to contaminated body fluids and aerosols. There are still many unknowns about the role that vertebrates play in the upkeep and spread of CCHFV. Critical information for our comprehension of CCHFV ecology may be gleaned through experimental studies conducted on wild animals and cattle. Therefore, the purpose of this review is to draw attention to current studies and publications that shed light on the intricate dynamic between CCHFV, vector, and host and demonstrate how biosurveillance at each stage aids in our knowledge of disease transmission, risk, and future research prospects.
Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) have the potential to provide an infinite source of tissues for regenerative medicine. Although defined xeno-free media have been developed, culture conditions for reliable propagation of hESCs still require considerable improvement. Here we show that recombinant E8 fragments of laminin isoforms (LM-E8s), which are the minimum fragments conferring integrin-binding activity, promote greater adhesion of hESCs and hiPSCs than do Matrigel and intact laminin isoforms. Furthermore, LM-E8s sustain long-term self-renewal of hESCs and hiPSCs in defined xeno-free media with dissociated cell passaging. We successfully maintained three hESC and two hiPSC lines on LM-E8s in three defined media for 10 passages. hESCs maintained high level expression of pluripotency markers, had a normal karyotype after 30 passages and could differentiate into all three germ layers. This culture system allows robust proliferation of hESCs and hiPSCs for therapeutic applications. The use of animal products as culture substrates for human embryonic stem cell and induced pluripotent stem cell culture raises numerous safety concerns in a therapeutic setting. Miyazaki et al.. show that minimal fragments of human laminins provide a more effective support for the culture of these cell types.