When and Why People Conceal Infectious Disease.

People sick with infectious illnesses face negative social outcomes, like exclusion, and may take steps to conceal their illnesses from others. In 10 studies of past, current, and projected illness, we examined the prevalence and predictors of infection concealment in adult samples of U.S. university students, health-care employees, and online crowdsourced workers (total N = 4,110). About 75% reported concealing illness in interpersonal interactions, possibly placing others in harm's way. Concealment motives were largely social (e.g., wanting to attend events like parties) and achievement oriented (e.g., completing work objectives). Disease characteristics, including potential harm and illness immediacy, also influenced concealment decisions. People imagining harmful (vs. mild) infections concealed illness less frequently, whereas participants who were actually sick concealed frequently regardless of illness harm, suggesting state-specific biases underlying concealment decisions. Disease concealment appears to be a widely prevalent behavior by which concealers trade off risks to others in favor of their own goals, creating potentially important public-health consequences.

A candidate competitive ELISA based on monoclonal antibody 3A8 for diagnosis of contagious bovine pleuropneumonia.

For the development of a competitive ELISA (cELISA) to detect serum antibodies against the Mycoplasma mycoides subsp. Mycoides (Mmm) (strain PG1), the causative agent of contagious bovine pleuropneumonia (CBPP), all the proteins of this pathogen were analyzed. Then, a specific extracellular region of a transmembrane protein with the potential for diagnosis was identified. After that, a monoclonal antibody (Mab) named 3A8 was obtained using this extracellular region as an immunogen. Finally, a cELISA was established with the extracellular domain of this transmembrane protein as the coating antigen, Mab 3A8 as the competitive antibody, and HRP-labeled goat anti-mouse IgG as the enzyme-labeled antibody. This established method was used to detect the antibody dynamic regularity of goats which are artificially immunized Mmm and was also compared with a commercial ELISA kit. Further, the sera of 1011 different cattle from border provinces of China were monitored using a candidate Mab 3A8 cELISA. The detection results of known background sera used in this study indicate that a candidate diagnostic marker was successfully identified by analyzing all the coding proteins of Mmm in this research, and the cELISA established based on the Mab 3A8 against this protein can detect CBPP-positive serum with specificity and has no cross-reaction with other related epidemic disease-positive sera. In addition, we tested the sera collected from the border areas of China using the established ELISA, and no positive sample was detected. The research protocol of the CBPP cELISA established in this study is different from the traditional method, which can greatly reduce the investment of manpower and capital and save development time. We believe that this study's protocol could serve as a reference for the development of detection methods for mycoplasma and other complex pathogens. KEY POINTS: • A Mmm-specific diagnostic marker was obtained based on protein characteristics. • A cELISA was established for CBPP serum antibody detection. • The serological investigation was conducted for CBPP in the border areas of China.

A molecular epidemiological investigation of contagious caprine pleuropneumonia in goats and captive Arabian sand gazelle (Gazella marica) in Oman.

BACKGROUND: Contagious caprine pleuropneumonia (CCPP) is a fatal WOAH-listed, respiratory disease in small ruminants with goats as primary hosts that is caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). Twelve CCPP outbreaks were investigated in 11 goat herds and a herd of captive Arabian sand gazelle (Gazella marica) in four Omani governorates by clinical pathological and molecular analysis to compare disease manifestation and Mccp genetic profiles in goats and wild ungulates. RESULTS: The CCPP forms in diseased and necropsied goats varied from peracute (5.8%), acute (79.2%) and chronic (4.5%) while all of the five necropsied gazelles showed the acute form based on the clinical picture, gross and histopathological evaluation. Colonies of Mccp were recovered from cultured pleural fluid, but not from lung tissue samples of one gazelle and nine goats and all the isolates were confirmed by Mccp-specific real time PCR. Whole genome-single nucleotide polymorphism (SNP) analysis was performed on the ten isolates sequenced in this study and twenty sequences retrieved from the Genbank database. The Mccp strains from Oman clustered all in phylogroup A together with strains from East Africa and one strain from Qatar. A low variability of around 125 SNPs was seen in the investigated Omani isolates from both goats and gazelles indicating mutual transmission of the pathogen between wildlife and goats. CONCLUSION: Recent outbreaks of CCPP in Northern Oman are caused by Mccp strains of the East African Phylogroup A which can infect goats and captive gazelles likewise. Therefore, wild and captive ungulates should be considered as reservoirs and included in CCPP surveillance measures.

Detection and genetic characterization of orf virus from sheep and goats in Nigeria.

Orf is a contagious, viral epitheliotropic disease of small ruminants. We investigated the molecular epidemiology of orf virus (ORFV) in breeds of small ruminants to determine the evolutionary diversity in Nigeria. Out of 54 small ruminants screened, the number of animals that were positive for ORFV in the three locations were 25. The distribution of positive animals by location were FCT 45.0% (n = 9/20), Oyo State 42.9% (6/14), and Plateau State 50.0% (n = 10/20). ORFV sequences from this study clustered with viruses detected in Taiwan, Iran, USA, and France. Our findings highlight the risk of transmission across geographic boundaries in Nigeria and West Africa, and reinforces the need for increased surveillance to prevent and control spread. Comprehensive characterization of ORFV in small ruminants as well as in humans in Nigeria is required to better elucidate the epidemiological dynamics and the virus evolution.

Mother-to-daughter transmission of hygienic anti-parasite behaviour in mandrills.

Social animals are particularly exposed to infectious diseases. Pathogen-driven selection pressures have thus favoured the evolution of behavioural adaptations to decrease transmission risk such as the avoidance of contagious individuals. Yet, such strategies deprive individuals of valuable social interactions, generating a cost-benefit trade-off between pathogen avoidance and social opportunities. Recent studies revealed that hosts differ in these behavioural defences, but the determinants driving such inter-individual variation remain understudied. Using 6 years of behavioural and parasite data on a large natural population of mandrills (Mandrillus sphinx), we showed that, when parasite prevalence was high in the population, females avoided grooming their conspecifics' peri-anal region (PAR), where contagious gastro-intestinal parasites accumulate. Females varied, however, in their propensity to avoid this risky body region: across years, some females consistently avoided grooming it, while others did not. Interestingly, hygienic females (i.e. those avoiding the PAR) were less parasitized than non-hygienic females. Finally, age, dominance rank and grooming frequency did not influence a female's hygiene, but both mother-daughter and maternal half-sisters exhibited similar hygienic levels, whereas paternal half-sisters and non-kin dyads did not, suggesting a social transmission of this behaviour. Our study emphasizes that the social inheritance of hygiene may structure behavioural resistance to pathogens in host populations with potential consequences on the dynamics of infectious diseases.

Contagious yawning in African painted dogs.

Contagious yawning (CY), which is yawning elicited by sensing another yawning, has been observed only in social species and is considered linked to high sociality. Although this idea-the social communication hypothesis-is supported by previous studies, investigating the occurrence of CY in various species remains necessary. We investigated the occurrence of CY in one of the most social canine species, the African painted dog (Lycaon pictus). We recorded 1387 yawn events from five pairs (10 individuals) in captivity. Temporal analysis showed that subsequent yawns occurred frequently within 15 s or 30 s after spontaneous yawns (SYs). SYs that were detectable by another individual (i.e., visible to the other individual or performed in close proximity) were more likely to elicit subsequent yawns by the other individual. However, the influence of contextual factors on CY differed according to the time window, implying that a conservative time window should be used to reduce overcounting of CY and misattribution of its determinants. The proportion of CY was positively related to time spent in physical proximity to another, a proxy for a social bond. Overall, our results provide the first evidence of CY in African painted dogs and further support the notion that such behavior is prevalent among social animals. They also strongly imply that an appropriate time window should be used to define yawn contagion.

Consistent Healthcare Safety Recommendation System for Preventing Contagious Disease Infections in Human Crowds.

The recent impact of COVID-19, as a contagious disease, led researchers to focus on designing and fabricating personal healthcare devices and systems. With the help of wearable sensors, sensing and communication technologies, and recommendation modules, personal healthcare systems were designed for ease of use. More specifically, personal healthcare systems were designed to provide recommendations for maintaining a safe distance and avoiding contagious disease spread after the COVID-19 pandemic. The personal recommendations are analyzed based on the wearable sensor signals and their consistency in sensing. This consistency varies with human movements or other activities that hike/cease the sensor values abruptly for a short period. Therefore, a consistency-focused recommendation system (CRS) for personal healthcare (PH) was designed in this research. The hardware sensing intervals for the system are calibrated per the conventional specifications from which abrupt changes can be observed. The changes are analyzed for their saturation and fluctuations observed from neighbors within the threshold distance. The saturation and fluctuation classifications are performed using random forest learning to differentiate the above data from the previously sensed healthy data. In this process, the saturated data and consistency data provide safety recommendations for the moving user. The consistency is verified for a series of intervals for the fluctuating sensed data. This alerts the user if the threshold distance for a contagious disease is violated. The proposed system was validated using a prototype model and experimental analysis through false rates, data analysis rates, and fluctuations.

How scared are Americans of the Zika virus? The role of threat, efficacy, and third-person perception to induce protective behaviors.

This study examines how public perception of threat and efficacy (on self and others) influence their tendency to take preventive action against the Zika virus by surveying 1,152 U.S. adults in Texas and Florida. Findings show that individuals were likely to take protective actions when they: (1) saw a high risk of the disease (high threat) and were confident about their ability to reduce the danger (high efficacy); and (2) perceived others as having a high risk (high threat), but lacked the ability to reduce the danger (low efficacy). Further, the study discusses practical implications for the design of public health campaigns.

Identification and molecular characterization of Orf virus infection in occupationally exposed women in South America.

Contagious Ecthyma (CE) is a severe exanthematous dermatitis caused by the Orf virus (ORFV) that mainly affects domestic small ruminants such as sheep and goats. It is a worldwide-distributed occupational zoonosis, particularly infecting those in close contact with animals or animal products such as shepherds, farmers and veterinarians, among others. In the present work, we report the first human CE case confirmed in Argentina. A phylogenetic analysis based on four gene sequences of the isolated strain responsible for the disease showed that this isolate grouped with other ORFV sequences that caused reported CE cases in sheep from the same Argentine province. We also sequenced a sample from a Chilean human case reported in 2017, whose phylogenetic analysis showed that it groups together with other Argentine isolates from locations close to the border with Chile.

Enhanced detection of Taylorella equigenitalis by qPCR using 'Dry' swabs.

Detection of Taylorella equigenitalis (CEMO) in the horse uses genital swabs. These swabs traditionally have been put in Amies charcoal transport medium for detection by culture but are also used for PCR. We determined the suitability of swabs without transport medium (Dry swabs) for CEMO PCR compared to swabs in Amies charcoal transport medium. The experiment was a factorial design using swab type and dilution of organism in culture suspensions, done in two parts. Simulated genital swabs were prepared in the laboratory by dipping in pairs into culture suspensions containing T. equigenitalis with or without other organisms, and then inserting them into a sleeve either with or without transport medium. In study 1, the difference in Ct value for the two swab types was compared. In study 2 genital swab material was then also added to culture suspensions and the swab types again compared. The swabs were tested by a validated quantitative PCR method. The Ct value of the PCR test was used as the measure for comparison, and the effect of variables assessed with linear regression. There was an 7.7% (6.5-8.9) higher mean Ct value of TM versus Dry swabs (P<0.001) overall. The Ct difference was more marked at higher dilutions. Addition of genital swab material had no effect on the Ct value. Dry swabs perform at least as well for PCR as swabs in Amies charcoal transport medium, especially when relatively low numbers of organism are present, and are advantageous for routine sampling when culture is not being used.

The Origin of Asexual Brine Shrimps.

AbstractDetermining how and how often asexual lineages emerge within sexual species is central to our understanding of sex-asex transitions and the long-term maintenance of sex. Asexuality can arise "by transmission" from an existing asexual lineage to a new one through different types of crosses. The occurrence of these crosses, cryptic sex, variations in ploidy, and recombination within asexuals greatly complicates the study of sex-asex transitions, as they preclude the use of standard phylogenetic methods and genetic distance metrics. In this study we show how to overcome these challenges by developing new approaches to investigate the origin of the various asexual lineages of the brine shrimp Artemia parthenogenetica. We use a large sample of asexuals, including all known polyploids, and their sexual relatives. We combine flow cytometry with mitochondrial and nuclear DNA data. We develop new genetic distance measures and methods to compare various scenarios describing the origin of the different lineages. We find that all diploid and polyploid A. parthenogenetica likely arose within the past 80,000 years through successive and nested hybridization events that involved backcrosses with different sexual species. All A. parthenogenetica have the same common ancestor and therefore likely carry the same asexuality gene(s) and reproduce by automixis. These findings radically change our view of sex-asex transitions in this group and show the importance of considering scenarios of asexuality by transmission. The methods developed are applicable to many other asexual taxa.

Host cell interactions of novel antigenic membrane proteins of Mycoplasma agalactiae.

BACKGROUND: Mycoplasma agalactiae is the main etiological agent of Contagious Agalactia syndrome of small ruminants notifiable to the World Organization for Animal Health. Despite serious economic losses, successful vaccines are unavailable, largely because its colonization and invasion factors are not well understood. This study evaluates the role of two recently identified antigenic proteins (MAG_1560, MAG_6130) and the cytadhesin P40 in pathogenicity related phenotypes. RESULTS: Adhesion to HeLa and sheep primary mammary stromal cells (MSC) was evaluated using ELISA, as well as in vitro adhesion assays on monolayer cell cultures. The results demonstrated MAG_6130 as a novel adhesin of M. agalactiae whose capacity to adhere to eukaryotic cells was significantly reduced by specific antiserum. Additionally, these proteins exhibited significant binding to plasminogen and extracellular matrix (ECM) proteins like lactoferrin, fibrinogen and fibronectin, a feature that could potentially support the pathogen in host colonization, tissue migration and immune evasion. Furthermore, these proteins played a detrimental role on the host cell proliferation and viability and were observed to activate pro-apoptotic genes indicating their involvement in cell death when eukaryotic cells were infected with M. agalactiae. CONCLUSIONS: To summarize, the hypothetical protein corresponding to MAG_6130 has not only been assigned novel adhesion functions but together with P40 it is demonstrated for the first time to bind to lactoferrin and ECM proteins thereby playing important roles in host colonization and pathogenicity.

Can contagious itch be affected by positive and negative suggestions?

Contagious itch can be evoked by observing people scratching. Verbal suggestions about to-be-received itch can influence itch intensity, as shown by placebo research, but it is unknown whether this extends to contagious itch. The current study aimed to replicate prior findings that listening to scratching and rubbing sounds elicits contagious itch, and to investigate whether suggestions can modulate this process. Healthy participants (n = 140) received positive or negative suggestions about itch in response to the sounds (aimed to decrease or increase expected itch, respectively), or no specific suggestions as a control. Participants listened to a number of audio fragments with scratching and rubbing sounds. The amount of expected itch as well as itch sensation after each audio fragment were measured by self-report. Suggestions had no effect on the expected itch. Both rubbing and scratching sounds significantly elicited itch in all groups. Scratching sounds induced more itch than rubbing sounds exclusively in the control group. These findings indicate that short suggestions might be not effective enough to modify the expectations of people regarding contagious itch. Furthermore, suggestions modulate contagious itch to some degree, but not in the hypothesized direction. Potential similarities and differences in the neurobiological mechanisms of contagious itch and nocebo effects are discussed.

Multi-locus sequence analysis reveals great genetic diversity among Mycoplasma capricolum subsp. capripneumoniae strains in Asia.

Multi-Locus Sequence Analysis (MLSA) of Mycoplasma capricolum subsp. capripneumoniae (Mccp) strains from Asia revealed unforeseen diversity and a central position for genotyping groups representing strains from Central/East Asia, suggesting a possible origin of contagious caprine pleuropneumonia in this continent. A better assessment of the emergence, diversity and distribution of Mccp in Asia and Africa calls for renewed efforts to dramatically enlarge the sample of strains. Availability and affordability in the field, added to superior typeability (directly from poor samples) and high stability, discriminatory power and concordance with epidemiological and phylogenetic analyses, make MLSA an excellent tool for such investigations.

Mycoplasma agalactiae ST35: a new sequence type with a minimal accessory genome primarily affecting goats.

BACKGROUND: Mycoplasma agalactiae, causing agent of contagious agalactia, infects domestic small ruminants such as sheep and goats but also wild Caprinae. M. agalactiae is highly contagious and transmitted through oral, respiratory, and mammary routes spreading rapidly in an infected herd. RESULTS: In an outbreak of contagious agalactia in a mixed herd of sheep and goats, 80% of the goats were affected displaying swollen udders and loss of milk production but no other symptom such as kerato-conjunctivitis, arthritis or pulmonary distress commonly associated to contagious agalactia. Surprisingly, none of the sheep grazing on a common pasture and belonging to the same farm as the goats were affected. Whole genome sequencing and analysis of M. agalactiae strain GrTh01 isolated from the outbreak, revealed a previously unknown sequence type, ST35, and a particularly small, genome size of 841'635 bp when compared to others available in public databases. Overall, GrTh01 displayed a reduced accessory genome, with repertoires of gene families encoding variable surface proteins involved in host-adhesion and variable antigenicity being scaled down. GrTh01 was also deprived of Integrative Conjugative Element or prophage, and had a single IS element, suggesting that GrTh01 has a limited capacity to adapt and evolve. CONCLUSIONS: The lack of most of the variable antigens and the Integrative Conjugative Element, both major virulence- and host specificity factors of a M. agalactiae strain isolated from an outbreak affecting particularly goats, indicates the implication of these factors in host specificity. Whole genome sequencing and full assembly of bacterial pathogens provides a most valuable tool for epidemiological and virulence studies of M. agalactiae without experimental infections.

Estimates of the collective immunity to COVID-19 derived from a stochastic cellular automaton based framework.

In the context of the propagation of infectious diseases, when a sufficient degree of immunisation is achieved within a population, the spread of the disease is ended or significantly decreased, leading to collective immunity, meaning the indirect protection given by immune individuals to susceptible individuals. Here we describe the estimates of the collective immunity to COVID-19 from a stochastic cellular automaton based model designed to emulate the spread of SARS-CoV-2 in a population of static individuals interacting only via a Moore neighbourhood of radius one, with a view to analyze the impact of initially immune individuals on the dynamics of COVID-19. This impact was measured by comparing a progression of initial immunity ratio-the percentage of immunised individuals before patient zero starts infecting its neighbourhood-from 0 to 95% of the initial population, with the number of susceptible individuals not contaminated, the peak value of active cases, the total number of deaths and the emulated pandemic duration in days. The influence of this range of immunities over the model was tested with different parameterisations regarding the uncertainties involved in the model such as the durations of the cellular automaton states, the contamination contributions of each state and the state transition probabilities. A collective immunity threshold of 55 % ± 2.5 % on average was obtained from this procedure, under four distinct parameterisations, which is in tune with the estimates of the currently available medical literature, even increasing the uncertainty of the input parameters.

Prevalence of bovine mastitis-related pathogens, identified by mass spectrometry in flies (Insecta, Diptera) captured in the milking environment.

Diptera (Insecta) are able to transmit approximately 200 pathogenic microorganisms to humans and animals, causing more than 65 diseases, including bovine mastitis, which constitutes a major cause of economic losses in the dairy industry. In this scenario, 217 adult specimens of Diptera were collected from nine farms and identified to the family and/or species level. Of the 11 families recorded, Muscidae was the most prevalent (152 out of 217; 70%). All Diptera specimens were subjected to microbiological culture using conventional and selective media, and isolates were then identified at the species level by mass spectrometry. In total, 275 microorganisms were identified, with a predominance of pathogens related to environmental bovine mastitis (166/275 = 60·4%), that is, Enterococcus species (70/275 = 25·4%) and Escherichia coli (49/275 = 17·8%). Nontraditional agents related to bovine mastitis (called miscellaneous) were detected in 28% (77/275), as well as microorganisms with well-known zoonotic behaviour (e.g. Bacillus cereus). This is study contributes with knowledge of diversity of microorganisms carried by Diptera in the dairy environment, including pathogens associated with environmental and contagious bovine mastitis, and agents with human relevance. To our knowledge, a three-part chromogenic selective medium used to microbial culture of milk on farms was used for the first time to identification of pathogens in Diptera.

The effect of fear of contagion/being contagious on depression, anxiety and stress levels of university students during the COVID-19 pandemic.

BACKGROUND: University students are generally quite active in social life. It is thought that their fear of contagion/being contagious will affect their depression, anxiety and stress levels due to the removal of restrictions and the increase in individual responsibilities. This study aims to determine the effect of fear of contagion/being contagious on depression, anxiety and stress levels of university students during the COVID-19 pandemic. METHODS: This study, descriptive and cross-sectional. Quantitative data were collected using the Demographic Information Form, the Fear of Contagion/Transmission Scale and the Depression-Anxiety-Stress Scale. The sample included 3500 participants. Descriptive statistics were used for the presentation of the results. The STROBE checklist for cross-sectional studies was used in this study. RESULTS: This study revealed that the depression, anxiety and stress levels of university students were above normal, and these results were mostly associated with females, those with a chronic disease and high fear of contagion/being contagious. CONCLUSION: The findings of this study may be useful for precautionary plans to support the psychological health of university students after the COVID-19 pandemic and in the fight against possible new pandemics. Considering the findings of our study, educational programs should be planned for youth in general and university students in particular to deal with depression, anxiety, stress and fears. RELEVANCE TO CLINICAL PRACTICE: The depression, anxiety and stress levels of university students were above normal. As the fear of contamination and contagion increases, the level of depression, anxiety and stress increases. Those with chronic diseases have high levels of depression, anxiety and stress. PATIENT OR PUBLIC CONTRIBUTION: Research data were collected through an online questionnaire. Data collection tools were prepared on Google Forms, and then, links were shared with student groups over social media. Students shared the link on their social platforms, allowing it to reach wider audiences.

A numerical evaluation of a novel recovery fresh air heat pump concept for a generic electric bus.

Since the outbreak of the worldwide COVID-19 pandemic, public transportation networks have faced unprecedented challenges and have looked for practical solutions to address the rising safety concerns. It is deemed that in confined spaces, operating heating units (and cooling) in non-re-circulation mode (i. e., all-fresh air mode) could reduce the airborne transmission of this infectious disease, by reducing the density of the pathogen and exposure time. However, this will expectedly increase the energy demand and reduce the driving range of electric buses. To tackle both the airborne transmission and energy efficiency issues, in this paper a novel recovery heat pump concept, operating in all-fresh air mode, was proposed. The novelty of this concept lies in its potential to be applied to already manufactured/in-service heat pump units as it does not require any additional components or need for redesigning the heating systems. In this concept, the cabin exhaust air is directed to pass through the evaporator of the heat pump system to recover part of the waste heat from the cabin and to improve the efficiency of the system. In this paper, a 0D/1D coupled model of a generic single-deck cabin and a heat pump system was developed in the Simulink environment of MATLAB (R2020b) software. The model was run in two different modes, namely the all-fresh air (as a baseline and a recovery heat pump concepts), and the air re-circulation mode (as a conventional heat pump concept with a 50% re-circulation ratio). The performance of these concepts was investigated to evaluate how an all-fresh air policy could affect the performance of the system, as well as the energy-saving potential of the proposed recovery concept. The performance of the system was studied under different ambient temperatures of -5 °C, 0 °C, and 5 °C, and for low and moderate occupancy levels. Results show that implementing the all-fresh air policy in the recovery and baseline concepts significantly improved the ventilation rate per person by at least 102% and at most 125%, compared to the air-re-circulating heat pump. Moreover, adopting the recovery concept reduced the power demand by at least 8% and at most 11%, compared to the baseline all-fresh air heat pump, for the selected fan and blower flow rates. The presented results in this paper along with the applicability of this concept to in-service mobile heat pumps could make it a feasible, practical, and quick trade-off solution to help the bus operators to protect people and improve the energy efficiency of their service.

Identification, molecular characterization, and pathological features of orf virus in sheep and goats in Punjab province, Pakistan.

Orf virus (ORFV) causes an acute, contagious, skin disease of sheep and goats which is economically important. The objectives of this study were to identify ORFV and to explore its pathological and phylogenetic profiles in 350 goats and 91 sheep of 14 districts of Punjab, Pakistan, from July 2020 to July 2021. Skin scrapings (total no. of samples = 441) of suspected animals were subjected to polymerase chain reactions, phylogenetic analysis, and pathological observations. The partial length of GIF/IL-2 gene (408 bp) was successfully amplified in 58/441 samples. Phylogenetic analysis of GIF/IL2 gene showed that the study isolates belonged to ORFV-cluster I, together with the viruses reported in India and China. Pakistan ORFV isolates were shared 97.6-98.7% nucleotide and 97.6-100% amino acid identities with the reference strain (NC_005336). Moreover, Chinese ORFV-isolates were detected unique multiple amino acid substitutions (F11L, Q21H, D27N, I46V, N49S, N82D, D103N, S129G) with study isolates. Naturally infected animals were anorexic, emaciated, dull, and depressed. The macroscopic lesions included multifocal to coalescing, ulceration followed by proliferative papules, pustules, and crust formation on the epidermis of gums, lips, mouth commissure, muzzles, nose, and udder. Histopathological examination revealed hyperplasia, anastomosing rete ridges formation and degenerative changes, including spongiosis and vacuolation of epidermal cells. Keratinocytes exhibited eosinophilic intracytoplasmic inclusion bodies with pyknotic and karyorrhexis nuclei. This is the first report on molecular characterization of ORFV from Pakistan, with insight into its pathogenesis and comparative analysis of pathological alterations and genetic diversity between ORFV strains reported in different geographical areas.