Implementing an Organizational Culture of Biosafety and Biosecurity in the SAP Institute.

An organizational culture of biosafety and biosecurity is critical for effective management of transboundary animal diseases. One essential aspect of this work is keeping important pathogens studied in veterinary laboratories under control. Türkiye is among the countries that are both endemic and disease-free for foot-and-mouth disease (FMD) virus, and it has a unique institute dedicated to FMD diagnosis, control, and vaccine production. To build an organizational safety culture within this institute and strengthen awareness of the importance of safe and secure handling of FMD, 4 staff members previously trained in biorisk management developed and provided trainings to all institute staff. The institute's 173 personnel were divided into 3 groups by job description based on direct or indirect work with FMD virus. All 3 groups received training that addressed biosecurity, biosafety, biorisk awareness, and insider threat; the trainings varied in length by group. Three-quarters (n=130, 75%) of all institute staff completed their training and were asked to complete knowledge surveys using a Likert scale survey before and after their training. A majority (n=104, 80%) of those participants completed both the pretraining and posttraining surveys. All 3 training groups' posttraining surveys showed improved awareness above baseline scores, and all 3 groups scores reached the targeted threshold goal. Group 2 demonstrated a realization that some of the knowledge and habits they had acquired through experience were incorrect. Scores for several individual questions decreased at posttraining, and these results will need further evaluation. The overall training results prompted the institute to provide periodic updates to employees to sustain the organizational safety culture. With this study, the institute now has a dedicated group of biorisk management representatives. This work serves as a wake-up call for established institutions that rely on staff experience to foster an organizational culture of biosafety and biosecurity.

Immune response against lumpy skin disease after simultaneous vaccination of cattle with sheep pox and goat pox and foot and mouth disease vaccines.

Foot and mouth disease (FMD) and Lumpy skin disease (LSD) are contagious viral diseases that cause significant economic damage in the livestock industry of countries. Cattle are vaccinated two times a year with FMD and sheep pox and goat pox vaccines (SGP) within 30-day intervals to combat both diseases in Türkiye. However, vaccinations in different periods increase vaccination costs, labor, and distress on animals. Therefore, it was aimed to determine the effects of simultaneous vaccination of FMD and SGP vaccines on the immunity against LSD and FMD in cattle. For this purpose, animals were divided into 4 groups; SGP vaccinated group (Group 1, n = 10), FMD vaccinated group (Group 2, n = 10), FMD and SGP simultaneously vaccinated group (Group 3, n = 10), and the unvaccinated control group (Group 4, n = 6). Blood samples were collected and analyzed to detect the antibody response against the LSD via Capripoxvirus (CaPV) ELISA and FMD by Virus Neutralisation test (VNT) and Liquid Phase Blocking ELISA (LPBE). A live virus challenge study was performed to determine the immune response against LSD. The mean antibody titers were determined protective levels on 28 days post vaccination (DPV) against FMDV serotypes O and A, respectively. The logarithmic difference of skin lesions was calculated log10 titer > 2.5. LSD genome could not be detected in the blood, eyes, and nose swap samples of the challenged animals on the 15th day via PCR. In conclusion, adequate protective immune response was provided against LSD when the SGP and FMD vaccines were used simultaneously in cattle.

Evaluation of Quil-A, E. coli DNA and Montanide™ ISA 206 adjuvant combination on the antibody response to foot-and-mouth disease vaccine in sheep.

Vaccination is one of the basic strategies in the fight against foot-and-mouth disease (FMD) in endemic regions. Today, commercially available FMD vaccines are prepared with inactive whole virion, which has low immunogenicity. Therefore, considerable effort has been devoted to finding novel adjuvants. Although mineral oils are among the most common adjuvants, it is still difficult to provide a long-term and robust immune response. Combined adjuvant systems are currently being studied to solve the problem. Saponins and CpG-ODNs have been shown to increase the immune response to vaccines individually in various studies. In this study, the effect of different adjuvants and their combinations (Quil-A, E. coli DNA, and MontanideTM ISA 206) on total and neutralizing antibody response in sheep was investigated. According to the results, the Quil-A group induced the highest antibody level, followed by the combination of Quil-A and the E. coli DNA group. The group containing E. coli DNA also caused a higher antibody response than the group containing only MontanideTM ISA 206 for certain days of sampling. These affordable alternatives of saponin and CpG sources can be used individually to increase the potency of the FMD vaccine for mass vaccinations of sheep. Keywords: foot-and-mouth disease; vaccine; adjuvant; Quil-A; E. coli DNA; combination of adjuvants.

The effects of simultaneous foot-and-mouth disease and Escherichia coli vaccination on the immunity of pregnant cows and their calves.

Fighting against infectious diseases with almost 18 million cattle spread over a wide geographical area is troublesome economically and physically. The administration of different vaccines simultaneously during the foot-and-mouth disease (FMD) vaccination campaign will help reduce both the vaccine stress of animals and the field veterinarians' workload. Diarrhea due to Escherichia coli (E. coli) is a significant problem and a major cause of mortality in calves, and thus, simultaneous application of FMD and E. coli vaccines to dams would prove efficient in field conditions. This study aimed to investigate the effect of simultaneous administration of four different locally produced gel and oil adjuvant inactivated E. coli vaccines with locally produced oil adjuvant inactivated FMD vaccine on neutralizing antibody levels and the properties of the antibodies in late-term pregnant dams and their calves. For this purpose, seven to eight-month-old pregnant dams (n = 146) were divided into two main groups to receive two doses (single and booster), or only single dose (no booster). FMD neutralizing antibody titers were evaluated by a virus neutralization test against serotype A, O, and Asia1. In addition, the properties of antibodies against serotype A were assessed by isotype and avidity ELISAs. E. coli antibodies were measured by the hemagglutination inhibition test. Results of the study revealed no safety problems in any dams after the vaccinations. On day 42, calves of the dams vaccinated simultaneously had higher neutralizing antibody titers against three serotypes than the calves of only FMD vaccinated dams. IgG1/IgG2 ratio was higher in single-dose groups than booster-dose groups against serotype A in calves. The avidity index was detected over the threshold value (24.5%). A positive correlation was found in the transmission of maternal antibodies from mothers to calves in simultaneous administration groups. In conclusion, during the FMD vaccine campaigns, FMD and E. coli vaccines can be simultaneously applied to dams in the 7th and 8th months of gestation. Results of this study has led to the inclusion of simultaneous application of FMD and E. coli vaccines in the 'Animal Disease and Animal Movement Control Program-2018' and these two vaccines have been applied in the field ever since as a solution for field veterinarians to save time and labor during vaccination.

Booster administration can make a difference in the antibody response to intradermal foot-and-mouth disease vaccination in cattle.

Foot-and-mouth disease (FMD) is a highly contagious and economically important viral disease of cloven-hoofed animals. Routine vaccination is one of the preferred methods of protection against this disease in endemic countries. For protective immunity against FMD, repeated immunizations with frequent administration are required. Intradermal immunization has many advantages over intramuscular administration of vaccines. In this study, a commercial tetravalent FMD vaccine adjuvanted with Montanide ISA 206 was administered to cattle via the intramuscular (2 mL [n = 10] and 0.5 mL [n = 9]) and intradermal (0.5 mL [n = 11]) routes. Booster doses were administered 28 days later using the same vaccine and routes. Serum samples were collected on days 0, 7, 14, and 28 post-vaccination (pv) and at 30 and 60 days post-booster. Homologous and heterologous virus neutralization tests and liquid-phase blocking and isotype ELISAs were used to measure the antibody response. The results showed that intradermal administration of quarter doses of the vaccine provides an equal or better virus neutralization antibody response than intramuscular administration of the same dose of vaccine after booster administration in cattle. This means that four times more cattle can be immunized with the same amount of vaccine using the intradermal route without compromising immunity.

Monitoring the performance of foot-and-mouth disease vaccines prepared against local strains in the face of antigenic evolution in the field.

National programs for foot-and-mouth disease (FMD) eradication includes the use of vaccination; Turkey which is endemic to FMD virus (FMDV) (except for the Thrace region) and there is a risk of incursion of exotic strains from eastern borders. In 2015, a devastating outbreak was caused by the A/ASIA/G-VII (G-VII) lineage, which led to the inclusion of a new vaccine strain (A/TUR/15) derived from this lineage in 3 months. Although most of the cattle population in Turkey was then immunized with A/TUR/15 (vaccine coverage: 92.8%), the G-VII lineage continued to cause outbreaks in the field despite the evidence of protection observed with A/TUR/15 in in vivo and in vitro tests. When G-VII field strains were examined, changes in their genomes were detected. As the lineage appeared to be evolving, an unconventional vaccination strategy was adapted which changed the vaccine strain with new variants of G-VII according to antigenic evolution. To assess the suitability of candidate vaccine strains derived from the variants of the G-VII lineage, three viral candidates were assessed (A/TUR/15, A/TUR/16 and A/TUR/17) by in vitro virus neutralization tests for r1 vaccine matching and in vivo heterologous challenge tests. Although all three vaccine strains were antigenically well matched with each other and other G-VII field viruses, due to continues outbreaks the vaccine strain was changed three times in 20 months from A/TUR/15 (Dec 2015) to A/TUR/16 (Dec 2016) and then to A/TUR/17 (Aug 2017). With this strategy serotype A has not been observed in the field since January 2018. This study highlights the importance of adapting the vaccine strains according to antigenic evolution as this could be a valuable combat strategy in endemic countries, rather than using well-known vaccine strain and relying only on the relationship coefficient (r1 ) value.

Determination of non-structural protein level for Turkey foot-and-mouth disease vaccine antigens during in-process.

PURPOSE: The success of foot-and-mouth disease (FMD) serological serosurveillance greatly depends on the FMD vaccine which does not include any non-structural proteins (NSPs) of the FMD virus. Since pure FMD vaccines from NSPs are used with the FMD eradication programs using DIVA (Differentiating Infected from Vaccinated Animals) tests. Apart from the in-vivo test defined in the World Organisation for Animal Health, two different test kits were developed in-process NSP detection purposes. The first test kit was developed in 2010 and the second one has been very recently developed in 2019. MATERIALS AND METHODS: In this study, the level of NSP has been examined by first-chemiluminescent filtration assisted (FAL)-enzyme-linked immunosorbent assay (ELISA) based in-vitro, in-process test kit for Turkey FMD vaccine antigen samples. A total of 94 samples were used. The critical maximum acceptable levels of NSP were determined after purification stage of samples. RESULTS: As a maximum NSP level, 70 ng NSP for the polyethylene glycol concentrated samples and 30 ng NSP for the vaccine antigen mixture samples were accepted. A mini repeatability study was also performed. The correlation between the NSP, total protein, and 146S particul quantity of samples were analyzed. CONCLUSION: As a conclusion, the chemiluminescent FAL-ELISA based test kit can be used for the NSP purity level determination of in-process samples.

Effect of simultaneous administration of foot-and-mouth disease (FMD) and anthrax vaccines on antibody response to FMD in sheep.

PURPOSE: Foot-and-mouth disease (FMD) and anthrax are important diseases in sheep. Vaccination is a favorable strategy against both infections. Simultaneous administration of vaccines does generally not impede the immune responses of each other, although there are some exceptions, and it may help reduce the labor and costs of vaccination as well as distress on animals. Although oil adjuvant FMD vaccine has been tried with live anthrax vaccine in cattle, there are no reports on the simultaneous use of both vaccines in sheep. MATERIALS AND METHODS: In this study, FMD seronegative sheep were used to investigate the impact of the simultaneous vaccination of FMD and anthrax on FMD antibody titers of sheep. Virus neutralization test and liquid phase blocking enzyme-linked immunosorbent assay were used to determine the antibody response to the FMD vaccine. RESULTS: The results demonstrated that both vaccines can be used simultaneously without any interference with the FMD response. Moreover, the simultaneous administration with anthrax vaccine had a stimulating effect on the early (day 7 post-vaccination) virus neutralization antibody response to the FMD vaccine. CONCLUSION: The simultaneous use of the FMD and anthrax vaccines did not hinder the response to the FMD vaccine in sheep.

Novel Approach to the Treatment of Diabetes: Embryonic Stem Cell and Insulin-Loaded Liposomes and Nanocochleates.

This study aims to investigate and compare the effects of insulin and embryonic stem-cell (ESC) loaded liposomes (LPs) and nanocochleate formulations and their PEGylated forms on the glucose levels. All formulations were characterized considering particle size, zeta potential, polydispersity index and encapsulation efficiencies. In-vitro insulin that releases from the formulations was determined using Franz-type diffusion cells. A cytotoxicity test revealed that none of the formulations was toxic to cells in any concentrations. The effects of the formulations on diabetic cells induced with glucose and streptozotocin (STZ) were then investigated in cell culture studies. Although glucose levels were decreased by the formulations after incubation, the liposomal formulations were found to be better. In experiments that were conducted on mice, it was observed again that blood glucose levels decreased successfully when diabetic pancreatic beta TC cells were incubated with the formulations, and all formulations were found to be effective in decreasing blood glucose levels in diabetic mice. Although ESC-loaded LPs were found to be the most effective formulation, LPs and nanocochleate formulations may also be used for the repair of pancreatic cells. This proposed ESC treatment is considered to be an attractive approach and a potential source for cell replacement therapy in the treatment of diabetes.

Influence of vaccine potency and booster administration of foot-and-mouth disease vaccines on the antibody response in calves with maternal antibodies.

Foot-and-mouth disease is one of the most important viral diseases of cloven-hoofed animals. Mass vaccination is an effective method to control the disease and is frequently utilized in endemic regions. Sufficient protection of young animals is important in mass vaccination campaigns. Maternal antibodies negatively affect the success of vaccination. Hence, determination of the optimal vaccination age is crucial for the uninterrupted protection of young animals. This study was performed to identify the effect of vaccine potency and booster administration on serum neutralizing antibody titers of calves with different levels of maternal antibodies. Calves (n = 111) on a state farm were used in this study. Oil adjuvant foot-and-mouth disease vaccines with 3 PD50 and 6 PD50 potencies were used with or without booster administration. Serum samples were collected each month up to day 120 postvaccination. Virus neutralization tests were used to measure the serum neutralizing antibody titers and estimate the protection period by using pre-determined cut-off values for protection. The results revealed that a vaccination with a 6 PD50 potency vaccine, preferably followed by a booster dose, should be used to overcome maternal immunity for incessant protection.

QS-21 enhances the early antibody response to oil adjuvant foot-and-mouth disease vaccine in cattle.

PURPOSE: One of the most important tools against foot-and-mouth disease, a highly contagious and variable viral disease of cloven-hoofed animals, is vaccination. However, the effectiveness of foot-and-mouth disease vaccines on slowing the spread of the disease is questionable. In contrast, high potency vaccines providing early protection may solve issues with the spread of the disease, escaping mutants, and persistency. To increase the potency of the vaccine, additives such as saponin and aluminium hydroxide are used. However, the use of saponin with an oil adjuvant is not common and is sometimes linked to toxicity. QS-21, which is less toxic than Quil A, has been presented as an alternative for use with saponin. In this study, the addition of QS-21 to a commercially available foot-and-mouth disease water-in-oil-in-water emulsion vaccine was evaluated in cattle. MATERIALS AND METHODS: After vaccination, serum samples were collected periodically over 3 months. Sera of the QS-21 and normal oil vaccine groups were compared via serum virus neutralization antibody titre and liquid phase blocking enzyme-linked immunosorbent assay antibody titre. RESULTS: The results showed that there was a significant early antibody increase in the QS-21 group. CONCLUSION: Strong early virus neutralizing antibody response will be useful for emergency or ring vaccinations against foot-and-mouth disease in target animals.

Chitosan-based systems for intranasal immunization against foot-and-mouth disease.

Inactivated conventional vaccines against foot-and-mouth disease (FMD) are used routinely in endemic countries and are effective against clinical disease. Increased systemic IgG levels can be obtained with these vaccines whereas local response at the mucosal sites where the virus primarily enters the organism and replicates remains very limited. The aim of this study was to develop a safe, non-invasive and antigen compatible system for mucosal delivery of the FMD antigen which induces both the systemic and local immunity. Gel formulations were prepared using different types of chitosan at different concentrations and were incorporated with the whole inactivated FMD virion. The immune responses in guinea pigs were determined following intranasal administration. Chitosan-based FMD vaccine formulations have been shown to induce FMD antigen-specific serum IgG and nasal IgA levels, the latter response being significantly stronger as compared to that obtained following subcutaneous administration of the FMD antigen in Freund's incomplete adjuvant. Our results suggest that intranasal immunization with inactivated FMD virion delivered in the presence of chitosan is very promising, inducing both systemic and local immune responses.

Prevalence, distribution, and host range of Peste des petits ruminants virus, Turkey.

Peste des petits ruminants virus (PPRV, genus Morbillivirus), which causes a severe disease in sheep and goats, has only recently been officially declared to be present in Turkey. We carried out a study to determine the prevalence, distribution, and host range of PPRV in Turkey. A total of 1,607 animals, reared in 18 different locations, were monitored for the presence of antibodies to PPRV and the related virus of large ruminants, Rinderpest virus (RPV). Only two farms had animals that were free of antibody responses to either disease. Prevalence for PPRV infection varied (range 0.87%-82.6%) and was higher in sheep (29.2%) than in goats (20%). The overall antibody responses to PPRV and RPV were 22.4% and 6.28%, respectively. Two PPRVs of lineage 4, which comprises many other PPRVs whose origins are in the Middle East, the Arabian Peninsula, and southern Asia, were isolated from Turkish sheep.