Gas chromatography-mass spectrometry method for determination ofβ-propiolactone in human inactivated rabies vaccine and its hydrolysis analysis

A simple method was established for the determination of β-propiolactone(BPL) in human inactivated rabies vaccine by gas chromatography-mass spectrometry(GC-MS). The determination was performed on an Agilent HP-INNOWAX(30 m ? 0.32 mm i.d., 0.25 mm) capillary column at the temperature of 80 °C.Electrospray ionization(ESI) was used by selective ion detection at m/z 42. The temperature for ESI source and inlet was set at 230 °C and 200 °C, respectively. Helium was used as the carrier gas at a flow rate of 25.1 m L/min. The total run time was 8 min. Acetonitrile and other components in the sample did not interfere with the determination of BPL. The results showed good linearity of BPL in the range of0.50–10.01 μg/mL, with the limit of detection and the limit of quantification of 0.015 μg/mL and0.050 μg/mL, respectively. Satisfactory precision was achieved for the current developed method. The method was applied to detect 6 batches of vaccine samples, and the results indicated that the target analyte BPL was present in three batches of unpurified samples, but was not detected in the purified samples, indicating the test samples were qualified. The established method was proved to be simple,versatile and sensitive, which can meet the requirements of quality control of BPL in human inactivated rabies vaccine.

Immunization Schedule of Liposomal Rabies Vaccine in Animals

The standard rabies vaccines recommended by WHO include Essen regimen, the Thai Red Cross two-site ID regimen and the eight-site ID regimen, and so on. The present schedules of rabies vaccine are all laborious and time consuming. We developed a new rabies vaccine with liposome as adjuvant（LipoRabV） and found that liposome could facilitate the inactivated rabies vaccine（RabV） to induce the more vigorous production of rabies virus neutralizing antibody（RVNA） in BALB/c mice and beagles. We established preliminary pre- and post-exposure prophylaxis schedules for LipoRabV. LipoRabV（0/14） could elicit similar RVNA level as RabV（0/7/28） by pre-exposure prophylaxis schedules in mice and beagles. LipoRabV（0/3/14） could elicit higher RVNA level vs. RabV（0/3/7/14/28） in BALB/c. The data indicate that the three-shot liposome-enhanced rabies vaccine could achieve a higher protection rate（survival rate 56.2%） by post-exposure prophylaxis compared with that of the RabV group（survival rate 40.6%） in mice. The data also indicate that the three-inoculation liposome-enhanced rabies vaccine could achieve a survival rate of 80.0% vs. RabV（70.0%） by post-exposure prophylaxis in beagles. The results show that the immunization schedule for LipoRabV could be preliminarily determined at 0 and 14 d for pre-exposure prophylaxis and at 0, 3 and 14 d for post-exposure prophylaxis.

Rabies DNA Vaccines: Current Status and Future

Rabies continues to be a significant cause of human and animal mortality, despite the availability of safe and effective prophylactics. Apart from limited access, the cost and complex schedules of rabies biologics often impact on the success of post-exposure prophylaxis in humans in the endemic countries. Mass vaccination of dogs, critical in rabies control, often fails to achieve its goal in rabies-endemic countries due to logistic, animal and vaccine-related issues. DNA vaccination has been proposed as a cheaper and efficient strategy for rabies prophylaxis, and its feasibility has been demonstrated in a number of animal models including companion animals, since 1994. Despite the proven efficacy, the technology suffers from a few drawbacks that limit its large-scale application, such as delayed and weaker immune responses in larger animals. Recent advances in the field of vector design and delivery hold promise for enhancement of rabies DNA vaccine efficacy. The present article provides an overview of developments in the field of DNA rabies vaccination and its future prospects.

IMT504: A New and Potent Adjuvant for Rabies Vaccines Permitting Significant Dose Sparing

Background: Rabies virus infection causes encephalitis, which is almost always fatal. Vaccination can be extremely effective at preventing disease but is prohibitively costly. Vaccine formulations allowing dose-sparing and fewer inoculations with faster antibody response would be extremely desirable. IMT504, an immunostimulatory non-CpG oligo-deoxynucleotide, is a highly potent vaccine adjuvant. Methods: Human and rat antibody measurements, and rat chal-lenge studies were performed. Results: In rats, highly effective immune responses with IMT504 were observed even after diluting vaccine up to 1/625. In highly lethal, live intracerebral rabies challenge studies, protection occurred even with extremely dilute vaccine plus IMT504. In humans, antibody titers developed faster and were significantly higher with IMT504-adjuvanted diluted vaccine vs non-adjuvanted vaccine (full strength or diluted). All five administered IMT504-adjuvanted diluted vaccine reached protective antibodies (≥0.5 IU/ml) after the second injection. After the third injection, individuals receiving IMT504-adjuvanted diluted vaccine reached levels approximately 10 times higher than controls (M ± SEM: 31.0 ± 10.9 vs 3.40 ± 0.99 IU/ml). Conclusions: These data suggest that IMT504 may allow fewer inoculations, highly significant dose-sparing of vaccine, rapid antibody production and protection from rabies. Extensive clinical studies are necessary to confirm if the use of IMT504 will permit significantly greater access to highly effective life-saving rabies vaccines.

Acute motor axonal neuropathy following anti-rabies human diploid cell vaccine:A rare case and review

Rationale:Guillain Barre syndrome(GBS)is an acute neurological illness leading to quadriparesis with respiratory involvement.It can be triggered by infections,vaccinations,surgery,trauma,transplantation and drugs.Anti-rabies cell culture vaccines introduced to overcome the high rate of neurological complications associated with tissue based rabies vaccine,can be very rarely associated with GBS.Patient concerns:A 50-year-old female presented with acute severe upper back pain evolving into pure motor quadriparesis following administration of human diploid cell vaccine for rabies.Diagnosis:Acute motor axonal neuropathy variant of GBS following anti-rabies human diploid cell vaccine.Interventions:Intravenous high dose steroids.Outcomes:Patient recovered completely within 1 month.Lessons:Although anti-rabies cell culture vaccines are highly immunogenic and safe,they are rarely associated with GBS.Clinicians should be aware of this link because prompt diagnosis and treatment can result in complete recovery and avoid complications.

Epidemic of Rabies and Effect of Its Vaccine against a Dog That Consecutively Attacked Ten People in One Day

On December 21, 20：10, a stray dog consecutively attacked 10 people in Lengshui Village, Ningyuan County, Yongzhou City, Hunan Province, China. The dog was killed by the local CDC staff and vicinity villager, its brain tissue sample was taken within 24 h. The epidemic focus was disinfected and the injured people received post exposure prophylaxis （PEP）. Pathogens were detected in the tissue sample by the provincial CDC. The immunity and safety of rabies vaccine were assayed after PEP, the injured people were regularly followed up in the following 2 y and 6 mon.

A Semi-quantitative Serological Method to Assess the Potency of Inactivated Rabies Vaccine for Veterinary Use

Potency is one of the most important indexes of inactivated vaccines.A number of methods have been established to assay the potency,of which the NIH test and single-dose mouse protection test are the "prescribed methods".Here,we report a method to semi-quantitatively assay the potency of an inactivated rabies vaccine,which uses fewer animals and takes less time to complete.Depending on the quality requirements of a vaccine(e.g.minimum potency),a rabies reference vaccine is,for example,diluted to the minimum potency,and 50 μL of the dilution is taken to inoculate 10 mice.The same amount of the test rabies vaccine is inoculated into another 10 mice.After two weeks,all mice are bled and serum samples are assayed for viral neutralizing antibody by the fluorescent antibody virus neutralization(FAVN) test.By comparing the median and interquartile range of antibody titers of the reference vaccine with those of the test vaccine,the test vaccine potency can be semi-quantitatively judged as to whether it is in accord with the required quality.The reliability of this method was also confirmed in dogs.The procedure can be recommended for batch potency testing during inactivated rabies vaccine production.

Breaking the Barriers to Access a Low Cost Intra-Dermal Rabies Vaccine Through Innovative “Pooling Strategy”

Background: In India every year an estimated 20,000 patients die of Rabies. Major reason for poor compliance to anti-rabies prophylaxis is the high cost of anti-rabies vaccine being prescribed intramuscularly (IM) as a routine i.e. 44.5 USD per course of five injections. In 1992 WHO recommended low cost intra-dermal rabies vaccination (IDRV), which costs only 7.5 USD or less per animal bite course. Methods: Interviews with doctors revealed that they were not prescribing intra-dermal anti rabies vaccination as they were either not aware or were not confident of this route of rabies vaccination. Also the vaccine vial did not have the label for “intra-dermal use”. These barriers were removed by advocacy efforts with policy makers & drug companies, credit sharing & team building, which led to starting of first intra dermal anti-rabies clinic of North India on 2nd August 2008. Results: Within a month of start of intra-dermal rabies vaccination clinic, i.e. by 2nd September, 2008, there was an increase in the hospital patient load by 2.8 times, and poor patients load by 3.2 times. In just less than two-year time, 200,000 USD of poor patients were saved and 5769 patients vaccinated. Each patient was asked to bring one vial on first visit & rest of doses were given “free” by pooling strategy. Pooling strategy involved distribution of one vial of vaccine among four persons and keep the three vials for use one by one by all the four patients on subsequent three visits. Another offshoot of the strategy was to prevent wasting of even few drops of vaccine that used to remain in each vial of 1 ml after distribution among four patients (0.2 mL or less). Out of more than 5000 vials utilised, every time we would transfer the left out drops of vaccine to the next new vial and use it immediately on a new pool of patients waiting for vaccination. We would, however, discard the unused vaccine after eight hours of reconstitution at the end of the day. The vaccine so saved turned to be a stock of more than 100 vials in less than two years that we were able to give free to more than 225 rag pickers, garbage collectors and newspaper hawkers on World Rabies Day, Sep 28, 2010. Conclusions: With intra-dermal clinic, we were able to successfully introduce the new cost effective intra-dermal method of rabies vaccination despite all odds & vested interests of companies & old mindset of doctors that had blocked this technique till now. This will go a long way in reducing the burden of disease & death due to rabies from India.

Pilot-Scale Production of Lyophilized Inactivated Rabies Vaccine Candidate in Vero Cells under Fully Animal Component-Free Conditions Using Microcarrier Technology and Laboratory Animal Trials

The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 - 12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 107 FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 - 4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study in Türkiye that used microcarriers.

Pup Vaccination Practices in India Leave People to the Risk of Rabies — Lessons from Investigation of Rabies Deaths Due to Scratch/Bite by Pups in Remote Hilly Villages of Himachal Pradesh, India

Rabies, a zoonotic disease, kills 55,000 persons every year globally and 20,000 persons in India. Two years back, we learnt of two deaths due to Rabies in remote village Shiv Shankar Garh of Arki block of District Solan and decided to investigate the deaths. Method: A rapid response team was constituted to investigate the deaths. We interviewed the villagers & family to conduct verbal autopsy. A line list of entire population of village and household contacts of the patients, who died, were made along with the line list of dogs and cattle. Results & Discussion: A-month-old stray pup brought home by the family and had caused an abrasion with its toes on the hands of both the deceased on June 2, 2011 while playing. The lady developed paralysis of the arm on July 3, 2011 and 3 days later developed symptoms of hydrophobia. She died on July 9, 2011. Her son had developed hydrophobia 10 days after that and died on July 19, 2011. Assumption that bite or abrasion by a small pup of one month cannot be fatal proved otherwise. Lack of awareness regarding the fatality of even a scratch and lack of knowledge regarding local treatment of the wound & vaccination of both human and pups, were the main reasons for the deaths. While such incidents keep on happening, and the veterinarians in India are refusing to vaccinate pups before three months of age, as pups may not develop immunity before that age, leaving unsuspecting people to the risk of rabies. Conclusions: Humans can be exposed to rabies even by pups below 3 months of age. Recommendation: Pup vaccination schedule in rabies endemic countries like India need revision. Veterinarians and public health experts need to strongly consider vaccinating pups at first contact with humans even if they are less than 3 months of age. A booster to the pup can be given at three months of age with subsequent yearly boosters.

Rabies in a postpandemic world:resilient reservoirs,redoubtable riposte,recurrent roadblocks,and resolute recidivism

Rabies is an ancient disease.Two centuries since Pasteur,fundamental progress occurred in virology,vaccinology,and diagnostics—and an understanding of pathobiology and epizootiology of rabies in testament to One Health—before common terminological coinage.Prevention,control,selective elimination,and even the unthinkable—occasional treatment—of this zoonosis dawned by the twenty-first century.However,in contrast to smallpox and rinderpest,eradication is a wishful misnomer applied to rabies,particularly post-COVID-19 pandemic.Reasons are minion.Polyhostality encompasses bats and mesocarnivores,but other mammals represent a diverse spectrum of potential hosts.While rabies virus is the classical member of the genus,other species of lyssaviruses also cause the disease.Some reservoirs remain cryptic.Although global,this viral encephalitis is untreatable and often ignored.As with other neglected diseases,laboratory-based surveillance falls short of the notifiable ideal,especially in lower-and middleincome countries.Calculation of actual burden defaults to a flux within broad health economic models.Competing priorities,lack of defined,long-term international donors,and shrinking local champions challenge human prophylaxis and mass dog vaccination toward targets of 2030 for even canine rabies impacts.For prevention,all licensed vaccines are delivered to the individual,whether parenteral or oral–essentially‘one and done’.Exploiting mammalian social behaviors,future‘spreadable vaccines’might increase the proportion of immunized hosts per unit effort.However,the release of replication-competent,genetically modified organisms selectively engineered to spread intentionally throughout a population raises significant biological,ethical,and regulatory issues in need of broader,transdisciplinary discourse.How this rather curious idea will evolve toward actual unconventional prevention,control,or elimination in the near term remains debatable.In the interim,more precise terminology and realistic expectations serve as the norm for diverse,collective constituents to maintain progress in the field.

Current Status of Canine Rabies in China

Abstract The number of human rabies cases acquired from dog bites constitutes a high proportion of the total rabies cases in China, although the number of human rabies cases has gradually decreased in recent years. The pivotal role of dogs in the spread of rabies indicates that controlling and preventing canine rabies could be a key step in eradicating human rabies in China. The primary aims of this review are to discuss the properties and pathogenesis of the rabies virus, the clinical signs and diagnosis of canine rabies, threshold host density and vaccination of dogs, and the prevention and control of canine rabies in China.

Establishment and Preliminary Application of a Rapid Fluorescent Focus Inhibition Test (RFFIT) for Rabies Virus

The World Health Organization (WHO) standard assay for determining levels of the rabies virus neutralization antibody (RVNA) is the rapid fluorescent focus inhibition test (RFFIT), which is used to evaluate the immunity effect after vaccination against rabies. For RFFIT, CVS-11 was used as the challenge virus, BSR cells as the adapted cells, and WHO rabies immunoglobulin (WHO STD) as the reference serum in this study. With reference to WHO and Pasteur RFFIT procedures, a micro-RFFIT procedure adapted to our laboratory was produced, and its specificity and reproducibility were tested. We tested levels of RVNA in human serum samples after immunization with different human rabies vaccines (domestic purified Vero cell rabies vaccine (PVRV) and imported purified chick embryo cell vaccine (PCECV)) using different regimens (Zagreb regimen and Essen regimen). We analyzed the levels of RVNA, and compared the immune efficacy of domestic PVRV and imported PCECV using different immunization regimens. The results showed that the immune efficacy of domestic PVRV using the Zagreb regimen was as good as that of the imported PCECV, but virus antibodies were generated more rapidly with the Zagreb regimen than with the Essen regimen. The RFFIT procedure established in our laboratory will enhance the comprehensive detection ability of institutions involved in rabies surveillance in China.

Immunizing Vulnerable Populations Like Rag Pickers, Garbage Collectors, Municipality Workers and Newspaper Hawkers against Rabies in Shimla Municipality, HP, India

Background: Rabies is a zoonotic disease and many vulnerable sections like rag pickers and municipality workers neglect animal bites due to ignorance of their potential deadly outcomes. Stray dogs abound in garbage pits and this population is exposed to their attacks. It should be a mandate for municipalities to help protect their sanitary workforce, especially rag pickers, from deadly infectious diseases such as Rabies, Hepatitis-B, HIV, Tetanus etc. Objectives: Objective of this study was to study methods to provide pre-exposure Rabies vaccination for such highly exposed populations by engaging them and understanding their perception of this disease through a constant dialogue with them. Methods: We started by engaging with the rag pickers to know how best to entice them to get themselves immunized. We then attempted to search literature for the most practical methods likely to succeed in reducing risk of rabies deaths in this population. Results: WHO approved 3 injections of 0.1 ml tissue culture vaccine on days 0, 7 and 21 were tried but were shown to result in many dropouts among rag pickers for repeat injections. We then followed a method where 0.1 ml of rabies vaccine was injected at 4 different anatomical sited in one setting. This proved acceptable and relatively inexpensive. A small number of subjects were studied by determination of neutralizing antibody by RFFIT, which proved immunogenic having anamnestic response on boosters given single IM or at 4 sites ID subsequently, implying that short schedule rabies pre-exposure vaccination can be done in high risk groups and may save lives if applied to the poorest that are highly exposed.

Causes of delay in offering rabies post-exposure prophylaxis services in Abadeh district of Iran

Objective:To identify the reasons for delayed reception of post-exposure prophylaxis(PEP).Methods:In this cross-sectional study,a total of 1407 individuals with animal bites who were referred to the Abadeh Rabies Treatment Center were investigated using the census method from January 2012 to December 2018.The patients were divided into two groups based on their delay times to referral and receive PEP:timely referral(less than 48 h after the bite)and delayed referral(equal to or longer than 48 h after the bite).Frequency,Chi-square,and logistic regression tests were used.Results:.The average delay time was(16.33±11.37)h.Low level of education(OR:3.87;95%CI:1.19-12.54;P=0.02),active economic age(21-35 and 36-50 years-old,OR:12.81;95%CI:3.16-51.97;P<0.001 and OR:3.83,95%CI:3.83-58.61;P<0.001 respectively),occupation(OR:9.16;95%CI:1.89-44.29;P=0.006),long distance from the rabies treatment center(OR:3.41;95%CI:2.03-5.72;P<0.001),bites by household and domestic animals(OR:12.22;95%CI:2.29-65.18,P=0.003),superficial injuries(OR:4.51;95%CI:1.38-14.73;P=0.01),and residence in rural area(OR:12.74;95%CI:6.58-24.66;P<0.001)had significant correlations with delayed referral of victims.Conclusions:To reduce the delay time,the high-risk groups should be informed about the importance of timely referral via educational measures.Furthermore,rabies treatment services should be rendered at the nearest possible center.

Exploring the Feasibility of a New Low Cost Intra-Dermal Pre &Post Exposure Rabies Prophylaxis Protocol in Domestic Bovine in Jawali Veterinary Hospital, District Kangra, Himachal Pradesh, India

Cattle are the backbone of household economy in rural areas of India and many of them die after bites by potentially rabid dogs, despite being given currently recommended five shots of intramuscular (IM) rabies vaccination as Post Exposure Prophylaxis (PEP). In 2016, seven of 21 bovine bitten by rabid dogs given IM rabies vaccination died due to rabies in Shimla Municipality. This scenario prompted the authors to look for a suitable protocol, based on human studies, to save animals. We tested various schedules of IDRV in bovine and found that a schedule of 0.2 ml given in middle 1/3rd of neck on day 0, 3, 7, 14 and 28 along with local wound infiltration of eRIG is sufficiently immunogenic and life saving in all of them, even if bitten by lab confirmed rabid dogs/mongoose as tested by CRI. Rabivac Vet, a Cell Culture Rabies Vaccine, available as 1 ml per vial was used off level for IDRV. While injecting the vaccine, a raised papule of ≥1 cm will appear slowly causing a peau d’orange appearance. All 60 bovine serum samples tested by RFFIT after IDRV, had titers more than 0.5 IU/ml on day 14. Thereafter, a total of 150 animals were given five doses of IDRV as PEP, with or without RIG, after their exposure to clinically or lab confirmed rabid dogs/mongoose and all survived for more than a year. Serum samples from 15 animals bitten by lab confirmed rabid dogs/mongoose were collected on day 14 and tested for RVNA by RFFIT from NIMHANS Bangalore and all had desired antibody titers above 0.5 IU/ml, without any immunosuppression. The RFFIT titers in 55% bovine in all groups were more than adequate after one year and 100% of them had anamnestic response to a single 0.2 ml booster given at one year. Few of the bovine and even one equine (Horse. Figure 4) brought for PEP at some of nearby vet hospitals were given IM rabies vaccine with local eRIG infiltration also survived. Local eRIG infiltration appeared to have covered the lacuna of longer window period required for indigenous antibodies production through IM route in bovine that are not sufficiently produced by day 14. While five times less vaccine was used in this low cost protocol and the survival was 100% compared to traditional IM protocol where survival was 66%. Pre-exposure prophylaxis was found to be effective as 0.2 ml dose of IDRV on day 0, 3, 7 and all bovine had titers higher than the desired by day seven after single 0.2 ml vaccine booster at one year. Our study points towards a possibility of having short schedules of three shots IDRV vaccination in bovine with or without local RIG (depending on presence or absence of wound/s) as PEP and single shot IDRV as PrEP, but further studies are required on a large number of animals. Our study also points out for allowing intra-dermal use in animals as well and labeling vaccines for the same as this is low cost more immunogenic and less painful compare to IM administration.

Interplay between rabies virus and the mammalian immune system

Rabies is a disease caused following infection of the brainby the rabies virus(RABV). The principle mechanism of transmission is through a bite wound. The virus infects peripheral nerves and moves to the central nervous system(CNS). There appears to be little involvement of other organ systems and little detectable immune stimulation prior to infection of the CNS. This failure of the mammalian immune system to respond to rabies virus infection leads, in the overwhelming majority of cases, to death of the host. To some extent, this failure is likely due to the exclusive replication of RABV in neurons and the limited ability to generate, sufficiently rapidly, an anti-viral antibody response in situ. This is reflected in the ability of post-exposure vaccination, when given early after infection, to prevent disease. The lack of immune stimulation during RABV infection preceding neural invasion is the Achilles heel of the immune response. Whilst many viruses infect the brain, causing encephalitis and neuronal deficit, none are as consistently fatal to the host as RABV. This is in part due to prior replication of many viruses in peripheral, non-neural tissue by other viruses that allows timely activation of the immune response before the host is overwhelmed. Our current understanding of the correlates of protection for rabies suggests that it is the action of neutralising antibodies that prevent infection and control spread of RABV. Furthermore, it tells us that the induction of immunity can protect and understanding how and why this happens is critical to controlling infection. However, the paradigm of antibody development suggests that antigen presentation overwhelmingly occurs in lymphoid tissue(germinal and non-germinal centres) and these are external to the CNS. In addition, the blood-brain-barrier may provide a block to the delivery of immune effectors(antibodies/plasma B-cells) entering where they are needed. Alternatively, there may be insufficient antigen exposure after natural infection to mount an effective response or the virus actively suppresses immune function. To improve our ability to treat this fatal infection it is imperative to understand how immunity to RABV develops and functions so that parameters of protectionare better defined.

Spontaneous Rabies in a Stray Bitch after Parturition Induced Immunosuppression —Investigating an Impending Outbreak of Rabies with One Health Approach

Background: Rabies is endemic in India and every half an hour a person dies of this dreaded disease. Stray roaming dogs, mostly unvaccinated, are most dangerous host in spread of rabies in India and in our state of Himachal Pradesh. Timely prophylaxis is the only method to save animal bite victims, including that of rabid dog bite patients. Objectives: Objective of this study was to investigate an impending outbreak of rabies in Shimla town in the absence of life saving rabies immunoglobulins (RIGs) in the market, and to know the source of infection by using one health approach and using epidemiological tools. Methods: On April 7, 2015, there was a sudden surge in cases of suspected rabid dog bites. Impending rabies outbreak was suspected as there were no RIGs available in the market. A rapid Response Team (RRT) consisting of the author, veterinary doctor, dog squad of Municipal Corporation (MC) Shimla along with the vehicles to impound rabid dogs was constituted to investigate the terror spread by two rabid dogs on biting spree in the Shimla Municipality. Results: A total of 18 people were bitten by suspected rabid dogs within three days period. A black bitch and a brown dog, on the identity of affected people, were captured by the dog squad of Municipality next day on April 8, 2015 and taken to Dog Sterilization Centre, Animal Birth Control (ABC) programme, MC Shimla for observation. The most furious Black bitch died of clinically confirmed symptoms of furious rabies after three days. The second rabid dog, brown in color, died after a month of observation due to paralytic dumb rabies. The brain of the brown dog was extracted for Fluorescent Antibody Testing (FAT) at central research Institute (CRI) Kasauli and was found to be positive for FAT. Follow up of patients was 100% by house to house visit and over telephone and no casualty was reported. Conclusion: While we were working on hypothesis of rabid dogs getting the infection from nearby forest about 8 - 10 KM away but on follow up of the patients, they reported that both the dog and bitch used to stay in the compound of their colony since the bitch was pregnant and there was no history of the bitch moving for away to forests neither any outside dog which was seen near them in the compound. This led us to think of other causes of what must had happened to the bitch that caused her to be rabid as for the past 9 years we had observed bitches becoming rabid after litter birth (Whelping) and making their pups rabid due to licking. Since the carrier state for rabies virus in bitches/dogs is known and a state of immunosuppression after whelping/parturition is also known, therefore there is a possibility of latent rabies virus getting activated due to immunosuppression after litter birth and it is thought to be as one of the probable causes of black bitch getting rabid and inflicting the infection to accompanying brown dog. We need to do further studies to ascertain this phenomenon before coming to a definite conclusion and suspect such a possibility in case a dam suddenly becomes rabid among a pack of stray dogs in rabies endemic countries like ours.

狂犬病暴露处置门诊接种人群的心理特点及护理干预措施研究

分析狂犬病暴露处置门诊接种人群的心理特点及护理干预措施。选取黔西市疾病预防控制中心狂犬病暴露处置门诊2022年3月—2022年4月接诊的325例狂犬病暴露接种者为研究对象,所有狂犬病暴露接种者都接受了健康教育、心理护理干预,对比心理护理干预前后狂犬病暴露接种者的焦虑评分、抑郁评分、医疗知识知晓率及接种疫苗依从率等指标。结果显示,干预后,狂犬病暴露接种者的焦虑及抑郁评分显著低于干预前且数据对比差异有统计学意义(P<0.05);干预后,狂犬病暴露接种者医疗知识知晓率、接种疫苗依从率都显著高于干预前且数据对比差异有统计学意义(P<0.05);干预后,狂犬病暴露接种者满意度显著高于干预前且数据对比差异有统计学意义(P<0.05)。研究发现,针对狂犬病暴露处置门诊接种人群的心理问题,对狂犬病暴露接种者进行针对性心理干预,可以缓解狂犬病暴露接种者负面情绪、提升其接种疫苗依从性。

Immunogenicity of multi-epitope-based vaccine candidates administered with the adjuvant Gp96 against rabies

Rabies, a zoonotic disease, causes > 55,000 human deaths globally and results in at least 500 million dollars in losses every year. The currently available rabies vaccines are mainly inactivated and attenuated vaccines, which have been linked with clinical diseases in animals. Thus, a rabies vaccine with high safety and efficacy is urgently needed. Peptide vaccines are known for their low cost, simple production procedures and high safety. Therefore, in this study, we examined the efficacy of multi-epitope-based vaccine candidates against rabies virus. The ability of various peptides to induce epitope-specific responses was examined, and the two peptides that possessed the highest antigenicity and conservation, i.e., AR16 and h PAB, were coated with adjuvant canineGp96 and used to prepare vaccines. The peptides were prepared as an emulsion of oil in water(O/W) to create three batches of bivalent vaccine products. The vaccine candidates possessed high safety. Virus neutralizing antibodies were detected on the day 14 after the first immunization in mice and beagles, reaching 5–6 IU/m L in mice and 7–9 IU/m L in beagles by day 28. The protective efficacy of the vaccine candidates was about 70%–80% in mice challenged by a virulent strain of rabies virus. Thus, a novel multi-epitope-based rabies vaccine with Gp96 as an adjuvant was developed and validated in mice and dogs. Our results suggest that synthetic peptides hold promise for the development of novel vaccines against rabies.