An index for multidimensional assessment of swine health.

Pork accounts for almost one-third of the meat consumed worldwide. Infectious diseases have a marked impact on pig production. Epidemiological indicators are considered the most useful criteria in decision-making; however, a health status assessment remains a challenge at the national and regional levels. This study proposes a health index including herd-losses, morbidity, fatality, and type of diseases, to rate the health situation in a region or country; it contributes to assessing the effectiveness of control, damage manifestation, and trends. It is a multidimensional index with a structure of triads and simple quantitative, semi-quantitative, and qualitative expressions that use flexible and dynamics limits. With it, we analyzed twenty-one countries in 2005-2018, focusing on African swine fever, classical swine fever, foot-mouth-disease, and porcine respiratory and reproductive syndrome, diseases that caused 72% of the morbidity. Our multidimensional approach estimates farm, local, and regional impact from infectious agents and outbreaks, and apprises trends aiming to be useful to control measures, strategic actions, and animal health policies.

FOOT-AND-MOUTH DISEASE IN EXPERIMENTALLY INFECTED MULE DEER (ODOCOILEUS HEMIONUS).

The only known outbreak of foot-and-mouth disease (FMD) in wildlife in the US occurred in mule deer (Odocoileus hemionus) in California in 1924-25. There is little recorded information on the pathogenesis and epidemiology of the disease in deer in that outbreak. In this experimental study, we compared the susceptibility of mule deer to FMD virus (FMDV) serotype O to that of cattle (Bos taurus). We also determined the potential for intra- and interspecies transmission of FMDV serotype O in mule deer and cattle, and assessed conventional laboratory tests in their ability to detect FMDV in mule deer. Two mule deer and one steer were each infected by intraepithelial tongue inoculation with 10,000 bovine tongue infective doses of FMDV, strain O1 Manisa. The inoculated steer and deer were kept in the same room with contact animals of both species. Exposed contact animals were moved to rooms with unexposed animals after becoming febrile. All mule deer (n=14) and cattle (n=6) developed clinical signs and lesions consistent with FMDV infection. Deer had a high prevalence of myocarditis and high mortality. Virus was transmitted between mule deer, from cattle to mule deer, and from mule deer to cattle. Virus and antibodies against nonstructural FMDV proteins in mule deer and cattle were detected by conventional laboratory tests. Virus shedding was detected by PCR and virus isolation up to 9 d postexposure in deer.

Genetic Determinants of Virulence between Two Foot-and-Mouth Disease Virus Isolates Which Caused Outbreaks of Differing Severity.

Individual foot-and-mouth disease virus (FMDV) strains reveal different degrees of infectivity and pathogenicity in host animals. The differences in severity among outbreaks might be ascribable to these differences in infectivity among FMDV strains. To investigate the molecular mechanisms underlying these differences, we estimated the infectivity of O/JPN/2000 and O/JPN/2010, which caused outbreaks of markedly different scales, in cell lines, Holstein cattle, and suckling mice. Viral growth of the two strains in cells was not remarkably different; however, O/JPN/2000 showed apparently low transmissibility in cattle. Mortality rates of suckling mice inoculated intraperitoneally with a 50% tissue culture infective dose (TCID50) of 10 for O/JPN/2000 and O/JPN/2010 also differed, at 0% and 100%, respectively. To identify genes responsible for this difference in infectivity, genetic regions of the full-length cDNA of O/JPN/2010 were replaced with corresponding fragments of O/JPN/2000. A total of eight recombinant viruses were successfully recovered, and suckling mice were intraperitoneally inoculated. Strikingly, recombinants having either VP1 or 3D derived from O/JPN/2000 showed 0% mortality in suckling mice, whereas other recombinants showed 100% mortality. This finding indicates that VP1, the outermost component of the virus particle, and 3D, an RNA-dependent RNA polymerase, are individually involved in the virulence of O/JPN/2010. Three-dimensional structural analysis of VP1 confirmed that amino acid differences between the two strains were located mainly at the domain interacting with the cellular receptor. On the other hand, measurement of their mutation frequencies demonstrated that O/JPN/2000 had higher replication fidelity than O/JPN/2010.IMPORTANCE Efforts to understand the universal mechanism of foot-and-mouth disease virus (FMDV) infection may be aided by knowledge of the molecular mechanisms which underlie differences in virulence beyond multiple topotypes and serotypes of FMDV. Here, we demonstrated independent genetic determinants of two FMDV isolates which have different transmissibility in cattle, namely, VP1 and 3D protein. Findings suggested that the selectivity of VP1 for host cell receptors and replication fidelity during replication were important individual factors in the induction of differences in virulence in the host as well as in the severity of outbreaks in the field. These findings will aid the development of safe live vaccines and antivirals which obstruct viral infection in natural hosts.

Outbreak investigations of foot and mouth disease virus in Nepal between 2010 and 2015 in the context of historical serotype occurrence.

Foot and Mouth Disease (FMD) is endemic in Nepal and causes substantial economic losses in the livestock industry. The goal of this study was to perform an epidemiological analysis of FMD outbreaks reported to the Veterinary Epidemiology Center, Tripureshwor, Nepal during 2010-2015, in order to strengthen the National FMD Control Program. These current data were considered in the context of historical data on FMD virus (FMDV) serotypes detected in the country between 1965 and 2015. During 2010-2015, a total of 1333 livestock holdings reported FMD outbreaks in Nepal. On average, 71.2 animals were affected in each outbreak, with a case fatality rate of 3.6%. FMD was reported throughout the country, and the proportion of affected holdings was not significantly among eco-zones, regions, or species. The Hill eco-zone had the highest number of holdings affected (782), followed by Mountain (304), and Terai (247). When analysed by the developmental region, the Western (381) and Central (368) Developmental Regions had the highest numbers of holdings affected. Cattle were the most frequently affected species (39%), followed by buffalo (33%), and goats (19%). FMD occurred throughout the year, with peaks in winter (December/January) and in the pre-monsoon period (April/May). Between 1965 and 2015 FMDV serotype O had the highest prevalence (81%), followed by Asia-1 (11%), A (6%), and C (2%). Serotype C was not detected after 1996, and only serotype O was reported after 2011. These descriptive analyses provide critical landmarks to establish baselines, and document early progress of the ongoing Progressive Control Pathway of FMD (PCP-FMD) which could be useful in Nepal and other South Asian nations.

Heterogeneity in a communal cattle-farming system in a zone endemic for foot and mouth disease in South Africa.

In South Africa, communal livestock farming is predominant in the foot and mouth disease control zone adjacent to the Greater Kruger National Park (KNP), where infected African buffaloes are common. During routine veterinary inspections of cattle in this area, a large amount of production and demographic parameters were being recorded. These data were collated for a five-year period (2003-2007) in three study sites to better understand the temporal dynamics and spatial heterogeneity in this system. A decreasing gradient from South to North with respect to both human and cattle population densities was observed. Rainfall and human population density alone could explain 71% of the variation in cattle density. Northern and central sites showed an overall decrease in total cattle numbers (15.1 and 2.9%, respectively), whereas a 28.6% increase was recorded in the South. The number of cattle owners in relation to cattle numbers remained stable during the study period. Only 4.0% of households in the South own cattle, compared to 13.7 and 12.7% in the North and Centre. The overall annual calving rate was 23.8%. Annual mortality rates ranged from 2.4 to 3.2%. Low calf mortality (2.1%) was recorded in the North compared to the South (11.6%). Annual off-take in the form of slaughter averaged 0.2, 11.7, and 11.0% in the North, Central and South sites, respectively. These figures provide valuable baseline data and demonstrate considerable spatial heterogeneity in cattle demography and production at this wildlife-livestock interface, which should be taken into consideration when performing disease risk assessments or designing disease control systems.

Induction of protection against foot-and-mouth disease virus in cell culture and transgenic suckling mice by miRNA targeting integrin αv receptor.

Foot-and-mouth disease virus (FMDV) is an RNA virus that causes a highly contagious disease in domestic and wild cloven-hoofed animals. Although vaccination has been used to protect animals against FMDV, there are shortcomings in the efficacy of the available vaccines. RNA interference (RNAi) is triggered by small RNA molecules, including short interfering RNAs and microRNAs (miRNAs), and the use of RNAi-based methods have demonstrated promise as an alternative method of controlling the transmission of FMDV. However, the method of delivery, short duration of siRNA and miRNA in vivo, and the genetic variability of FMDV confound the use of RNAi-based strategies for FMDV control. FMDV has been shown to exploit host-cell integrins as cell-surface receptors to initiate infection. We selected the gene for the integrin αv subunit as an RNAi target, and constructed three αv-specific miRNA expression plasmids. The effects of these miRNAs on FMDV infection were examined in PK-15 cells and transgenic suckling mice. In PK-15 cells, the expression of the αv-specific miRNAs significantly inhibited the expression of integrin αv receptor and decreased FMDV infection. The transgenic mice were generated by integrating the αv-specific miRNA expression cassette using pronuclear microinjection. When challenged with a dose of FMDV ten times greater than the LD50, the survival rate of transgenic suckling mice was approximately six-fold higher than that of their non-transgenic littermates, indicating that the interference of the miRNAs significantly reduced FMDV infection in the transgenic mice. This is the first report of limiting FMDV attachment to cellular receptors using miRNA-mediated gene knock down of cell-surface receptors to significantly reduce FMDV infection in cell culture and transgenic suckling mice.

A recombinant adenovirus bicistronically expressing porcine interferon-α and interferon-γ enhances antiviral effects against foot-and-mouth disease virus.

Foot-and-mouth disease (FMD) is a virulent and economically costly disease in domestic livestock. Since the current vaccine available against FMD provides no protection until 7days postvaccination, the only alternative method to halt the spread of the FMD virus (FMDV) during outbreaks is by the application of anti-viral agents. The combination of recombinant adenovirus expressing type I interferon (IFN-α) and adenovirus expressing type II IFN (IFN-γ) has been reported to be an effective anti-viral treatment strategy against FMDV. Nevertheless, the recombinant adenovirus mixture may be inefficient because of the low anti-viral efficiency of IFN-γ compared to that of IFN-α. In this study, we generated a recombinant adenovirus co-expressing porcine IFN-α and IFN-γ in tandem using an FMDV 2A sequence to mediate effective cleavage of the two proteins (referred to as Ad-porcine IFN-αγ). We demonstrated that both recombinant porcine IFN-α and IFN-γ were expressed and interferon stimulated gene (ISG)s related with IFN-α and IFN-γ were induced in porcine kidney (IBRS-2) cells infected with Ad-porcine IFN-αγ. Additionally, the anti-viral effects of Ad-porcine IFN-αγ against FMDV were enhanced both in IBRS-2 cells and in CD-1 (ICR) suckling mice compared to that of adenovirus expressing only a single protein. We propose that Ad-porcine IFN-αγ could be a rapid, highly efficient, convenient anti-viral agent against FMDV.

In vitro maturation and fertilization in the Nilgai (Boselaphus tragocamelus) using oocytes and spermatozoa recovered post-mortem from animals that had died because of foot and mouth disease outbreak.

The ability to rescue gametes from endangered or wildlife species and to subsequently produce viable embryos holds tremendous potential as a means to increase the population size of endangered or wildlife species. The objective of this study was to assess the developmental competence of gametes recovered from nilgai that had died because of foot and mouth disease outbreak. Oocytes collected from the ovaries of seven dead nilgais were allowed to mature in vitro and were tested for developmental potential by in vitro fertilization (IVF) with epididymal spermatozoa collected also post-mortem. The average number of oocytes (n = 517) recovered per ovary was 36.9, and the side (right or left), size and weight of the ovaries had no significant effect on the number and quality of oocytes recovered. In vitro maturation studies indicated that the proportion of matured oocytes (MII stage) at 18, 24 and 30 h was 55.6%, 63.4% and 63.6%, respectively. Furthermore, 43% of the matured oocytes cleaved following in vitro fertilization and 12% of the cleaved oocytes (6/49) developed to the 4-8 cell stage. These findings suggest that the gametes recovered from nilgai post-mortem could be utilized for in vitro production of embryos.

Development of a foot-and-mouth disease infection model in severe combined immunodeficient mice for the preliminary evaluation of antiviral drugs.

Recent European guidelines facilitate the use of emergency vaccines during outbreaks of foot-and-mouth disease. Antiviral drugs could be used as a complementary measure. This study aimed at developing a small animal model to assess the in vivo activity of early antiviral lead molecules with anti-foot-and-mouth disease virus (FMDV) activity in vitro. In a first attempt, several FMDV strains were titrated in Balb/c mice. Inoculations with O1 Manisa or C1 Noville did not induce clinical disease, whereas Asia1 Shamir induced death too rapidly [i.e. within 4 days post-inoculation (dpi)]. Therefore, we switched to severe combined immunodeficient mice which are frequently used as a model for viral infections and experimental therapeutics. Strain O1 Manisa did not induce clinical disease, but titrations with A22 Iraq, C1 Noville or Asia1 Shamir resulted in virus-induced morbidity (including respiratory problems and weight loss) with subsequent mortality. Inoculations with strain A22 Iraq resulted in a reproducible mean time of death of 6 dpi (this was shorter for the other strains). In this newly developed rodent model, strain A22 Iraq seems the most suited to assess the in vivo anti-FMDV activity of selective inhibitors of FMDV.

Chitosan can stop or postpone the death of the suckling mice challenged with foot-and-mouth disease virus.

UNLABELLED: In the study, a method called "hardening in liquid phase" for preparing chitosan granules with glutaraldehyde as crosslinker and Tween 80 as surfactant and paraffin liquid as dispersant was established. The chitosan granules were light yellow and insoluble in water or oil, but they swelled in acid solution and narrowed in neutral or alkaline solution. Furthermore, some of characteristics of the chitosan granules were revealed. (a) Stability: Their shapes were stable at pH 7.0 and pH 8.0 and -30 degrees C-120 degrees C. The shelf life is at least one year in vitro at room temperature. (b) SAFETY: Some experiments of their lethal effect to suckling mice and pathogenicity to mature mice proved the chitosan granules were harmless. (c) Antiviral activity: Some suckling mice injected with chitosan granules were still alive or delayed death compared with control group when they challenged with foot-and-mouth disease virus (FMDV). Such anti-FMDV capacity could maintain 1 week and was the strongest on the third day.

Inhibition of foot-and-mouth disease virus replication in vitro and in vivo by small interfering RNA.

By using bioinformatics computer programs, all foot-and-mouth disease virus (FMDV) genome sequences in public-domain databases were analyzed. Based on the results of homology analysis, 2 specific small interfering RNA (siRNA) targeting homogenous 3D and 2B1 regions of 7 serotypes of FMDV were prepared and 2 siRNA-expression vectors, pSi-FMD2 and pSi-FMD3, were constructed. The siRNA-expressing vectors were used to test the ability of siRNAs to inhibit virus replication in baby hamster kidney (BHK-21) cells and suckling mice, a commonly used small animal model. The results demonstrated that transfection of BHK-21 cells with siRNA-expressing plasmids significantly weakened the cytopathic effect (CPE). Moreover, BHK-21 cells transiently transfected with short hairpin RNA (shRNA)-expressing plasmids were specifically resistant to the infection of the FMDV serotypes A, O, and Asia I and this the antiviral effects persisted for almost 48 hours. We measured the viral titers, the 50% tissue culture infective dose (TCID50) in cells transfected with anti-FMDV siRNAs was found to be lower than that of the control cells. Furthermore, subcutaneous injection of siRNA-expressing plasmids in the neck of the suckling mice made them less susceptible to infection with O, and Asia I serotypes of FMDV.

Maasai perception of the impact and incidence of malignant catarrhal fever (MCF) in southern Kenya.

We investigated the perceived impact of malignant catarrhal fever (MCF) to pastoralists in Isinya Division, a wildlife dispersal area of Nairobi National Park, and used a range of participatory epidemiology methodologies. We compared the relative importance, incidence and impact of MCF compared to other locally defined important diseases with a total of 158 respondents in 11 group meetings and 21 household meetings in July 2004. Direct losses due to disease were investigated through lowered prices as a result of the emergency sale of disease-infected animals. Overall, Maasai in Isinya Division perceived east coast fever (ECF) to be the most important cattle disease and to have the highest incidence. Anthrax was considered to have the largest impact. In areas within or adjacent to the wildebeest calving zone, MCF was perceived to be the most important cattle disease and also to have the largest impact. Outside the calving zone, MCF was considered the fourth-most important disease with the fourth largest impact, and these were areas where wildebeest were less common. MCF was also the fourth-most common disease, and across the Division incidence was estimated at 5% in calves and 10% in adults. However, MCF incidence varied greatly throughout the study area, from 3% to 12%, and the highest incidence risks were found in areas where wildebeest came to calve. The percent drop in sale price per animal infected with MCF was estimated at 50% for MCF for the year 2003-2004. Forced avoidance movements away from wildebeest calves were reported to decrease livestock production due to loss of access to prime grazing sites. As suggested by pastoralists in this study, the development of compensation schemes or incentives from wildlife would reduce the conflict between livestock keeping and wildlife conservation.

Foot-and-mouth disease virus (FMDV) causes an acute disease that can be lethal for adult laboratory mice.

Foot-and-mouth disease virus (FMDV) is a picornavirus that causes an acute vesicular disease of cloven-hoofed animals. This virus continues to be threat to livestock worldwide with outbreaks causing severe economic losses. However, very little is known about FMDV pathogenesis, partially due to the inconveniences of working with cattle and swine, the main natural hosts of the virus. Here we demonstrate that C57BL/6 and BALB/C adult mice are highly susceptible to FMDV infection when the virus is administered subcutaneously or intraperitoneally. The first clinical signs are ruffled fur, apathy, humped posture, and wasting, which are followed by neurological signs such as hind-limb paralysis. Within 2-3 days of disease onset, the animals die. Virus is found in all major organs, indicating a systemic infection. Mice developed microvesicles near the basal layer of the epithelium, event that precedes the vesiculation characteristics of FMD. In addition, a lymphoid depletion in spleen and thymus and severe lymphopenia is observed in the infected mice. When these mice were immunized with conventional inactivated FMDV vaccine, they were protected (100% of vaccinated animals) against challenge with a lethal dose of FMDV. The data indicate that this mouse model may facilitate the study of FMDV pathogenesis, and the development of new effective vaccines for FMD.

[Culling of animals in large herds, experiences from the Netherlands]. / Keulung von Tieren in Grossbeständen, Erfahrungen in den Niederlanden.

The outbreaks of classical swine fever in 1997-1998 and foot- and mouth-disease in 2001 provided a lot of experiences in the culling of animals. These experiences, as well as the aspects of animal welfare and public acceptance are described. In the future these experiences will help to carry out culling in a more efficient way including improved aspects of animal welfare and public acceptance.