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1.  Alternative medicine: methinks the doctor protests too much and incidentally befuddles the debate. 
Journal of Medical Ethics  1992;18(1):23-25.
Dr Kottow in his paper Classical medicine v alternative medical practices (1) places the alternative/orthodox medicine debate within an historical context of anti-quackery literature. My paper explores the nature of science as it is applied to clinical practice and challenges the narrow view of the diagnostic process as outlined by Dr Kottow. Research methodologies more appropriate to 'whole person' medicine are suggested as having more ethical value than those based on the clinical trial.
PMCID: PMC1376080  PMID: 1573645
2.  ‘Saving the lives of our dogs’: the development of canine distemper vaccine in interwar Britain 
This paper examines the successful campaign in Britain to develop canine distemper vaccine between 1922 and 1933. The campaign mobilized disparate groups around the common cause of using modern science to save the nation's dogs from a deadly disease. Spearheaded by landed patricians associated with the country journal The Field, and funded by dog owners and associations, it relied on collaborations with veterinary professionals, government scientists, the Medical Research Council (MRC) and the commercial pharmaceutical house the Burroughs Wellcome Company (BWC). The social organization of the campaign reveals a number of important, yet previously unexplored, features of interwar science and medicine in Britain. It depended on a patronage system that drew upon a large base of influential benefactors and public subscriptions. Coordinated by the Field Distemper Fund, this system was characterized by close relationships between landed elites and their social networks with senior science administrators and researchers. Relations between experts and non-experts were crucial, with high levels of public engagement in all aspects of research and vaccine development. At the same time, experimental and commercial research supported under the campaign saw dynamic interactions between animal and human medicine, which shaped the organization of the MRC's research programme and demonstrated the value of close collaboration between veterinary and medical science, with the dog as a shared object and resource. Finally, the campaign made possible the translation of ‘laboratory’ findings into field conditions and commercial products. Rather than a unidirectional process, translation involved negotiations over the very boundaries of the ‘laboratory’ and the ‘field’, and what constituted a viable vaccine. This paper suggests that historians reconsider standard historical accounts of the nature of patronage, the role of animals, and the interests of landed elites in interwar British science and medicine.
doi:10.1017/S0007087413000344
PMCID: PMC4014013  PMID: 24941736
3.  Neither consenting nor protesting: an ethical analysis of a man with autism 
Journal of Medical Ethics  2000;26(4):277-281.
This article critically examines the 25 June 1998 decision by the House of Lords regarding the psychiatric admission of a man with autism.1 Mr L was able neither to consent to, nor refuse, that admission and the disposition of his case illuminates the current debate regarding best interests of vulnerable adults by the judiciary and the psychiatric profession. This article begins with the assumption that hospitalisation was not the optimum response to Mr L's condition, provides alternative approaches to the interpretation of best interest and examines principles of liberty, anti-discrimination, and equal protection.
Key Words: Informal admission • autism • discrimination
doi:10.1136/jme.26.4.277
PMCID: PMC1733269  PMID: 10951924
4.  Protest of doctors: a basic human right or an ethical dilemma 
BMC Medical Ethics  2014;15:24.
Background
Peaceful protests and strikes are a basic human right as stated in the United Nations’ universal declaration on human rights. But for doctors, their proximity to life and death and the social contract between a doctor and a patient are stated as the reasons why doctors are valued more than the ordinary beings. In Pakistan, strikes by doctors were carried out to protest against lack of service structure, security and low pay. This paper discusses the moral and ethical concerns pertaining to the strikes by medical doctors in the context of Pakistan. The author has carefully tried to balance the discussion about moral repercussions of strikes on patients versus the circumstances of doctors working in public sector hospitals of a developing country that may lead to strikes.
Discussion
Doctors are envisaged as highly respectable due to their direct link with human lives. Under Hippocrates oath, care of the patient is a contractual obligation for the doctors and is superior to all other responsibilities. From utilitarian perspective, doctors’ strikes are justifiable only if there is evidence of long term benefits to the doctors, patients and an improvement in service delivery. Despite that, it is hard to justify such benefits against the risks to the patients. Harms that may incur to the patients include: prolongation of sufferings, irreversible damage to health, delay in treatment, death, loss of work and waste of financial resources.
In a system of socialized medicine, government owing to greater control over resources and important managerial decisions should assume greater responsibility and do justice to all stakeholders including doctors as well as patients. If a doctor is underpaid, has limited options for career growth and is forced to work excessively, then not only quality of medical care and ability to act in the best interests of patients is adversely affected, it may also lead to brain drain.
Summary
There is no single best answer against or in favor of doctors’ industrial action. The author calls for the debate and discussion to revitalize the understanding of the ethical predicaments of doctors’ strikes with patient care as the priority.
doi:10.1186/1472-6939-15-24
PMCID: PMC3995928  PMID: 24612947
Ethical dilemma; Doctors’ strikes; Pakistan; Service structure; Lack of security; Low pay; Socialized medicine; Utilitarian theory; Brain drain
5.  SUBCLINICAL INFECTION OF DOGS BY CANINE-ADAPTED MEASLES VIRUS EVIDENCED BY THEIR SUBSEQUENT IMMUNITY TO CANINE DISTEMPER VIRUS 
Journal of Bacteriology  1961;82(5):702-705.
Moura, Roberto A. (Chas. Pfizer and Company, Inc., Terre Haute, Ind.) and Joel Warren. Subclinical infection of dogs by canine-adapted measles virus evidenced by their subsequent immunity to canine distemper virus. J. Bacteriol. 82:702–705. 1961.—Young dogs were inoculated with virulent measles virus which had been adapted to canine kidney or human amnion cell culture. None of the animals showed any clinical symptoms nor could virus be isolated from the blood, although measles-neutralizing and complement-fixing antibodies developed during convalescence. All dogs failed to develop antibody to canine distemper. However, when these and normal control animals were subsequently inoculated intracerebrally with virulent distemper virus, each of the controls succumbed to typical symptoms, whereas all of the measles-immune dogs survived. These results suggest that the cross-protection conferred by measles against canine distemper virus infection involves factors other than humoral antibody. The immunity persists for a considerable length of time.
PMCID: PMC279238  PMID: 14476677
6.  Protection against canine distemper virus in dogs after immunization with isolated fusion protein. 
Journal of Virology  1986;58(2):536-541.
Canine distemper virus attachment (hemagglutinin [H] equivalent) and fusion (F) antigens were purified by affinity chromatography with monoclonal antibodies. The purified antigens were used to immunize groups of three dogs. Radioimmune precipitation assays with sera from these animals showed that the F antigen preparation was pure and induced only an F polypeptide-specific antibody response but that the H antigen preparation had a slight contamination by the F antigen. Immunized animals were challenged with virulent canine distemper virus. Two animals in each group developed pronounced humoral and cellular immune responses after challenge. Among these infected animals, only the dogs immunized with H antigen developed symptoms, albeit mild. In contrast, three nonimmunized control animals developed severe disease, with a fatal outcome in two cases. The complete resistance against challenge in two dogs was interpreted to reflect in one case anti-F immunity and in the other case most likely a high level of anti-H immunity. It is suggested that the F antigen may be of particular interest for the development of morbillivirus and possibly other paramyxovirus subunit or synthetic vaccines, because it can induce immunity capable of blocking virus infection and in situations of virus replication prevent the emergence of symptoms.
Images
PMCID: PMC252942  PMID: 3754590
9.  Rapid and Sensitive Detection of Immunoglobulin M (IgM) and IgG Antibodies against Canine Distemper Virus by a New Recombinant Nucleocapsid Protein-Based Enzyme-Linked Immunosorbent Assay 
Journal of Clinical Microbiology  1999;37(4):1049-1056.
Canine distemper morbillivirus (CDV) infection causes a frequently fatal systemic disease in a broad range of carnivore species, including domestic dogs. In CDV infection, classical serology provides data of diagnostic and prognostic values (kinetics of seroconversion) and is also used to predict the optimal vaccination age of pups. Routine CDV serology is still based on time- and cost-intensive virus neutralization assays (V-NA). Here, we describe a new capture-sandwich enzyme-linked immunosorbent assay (ELISA) that uses recombinant baculovirus-expressed nucleocapsid (N) protein of a recent CDV wild-type isolate (2544/Han95) for the detection of CDV-specific antibodies in canine sera. Recombinant antigen was produced with high efficacy in Heliothis virescens larvae. The capture-sandwich ELISA enabled a clear-cut qualitative evaluation of the CDV-specific immunoglobulin G (IgG) and IgM serostatuses of 196 and 35 dog sera, respectively. Inter-rater agreement analysis (κ = 0.988) indicated that the ELISA can be used unrestrictedly as a substitute for the V-NA for the qualitative determination of CDV-specific IgG serostatus. In an attempt to semiquantify N-specific antibodies, a one-step-dilution (alpha method) IgG-specific ELISA was implemented. Alpha values of ≥50% showed very good inter-rater agreement (κ = 0.968) with V-NA titers of ≥1/100 50% neutralizing dose (ND50) as measured against the central European CDV wild-type isolate 2544/Han95 in canine sera originating from northern Germany. An ND50 titer of 1/100 is considered a threshold, and titers of ≥1/100 indicate a resilient, protective immunity. CDV N-specific antibodies of the IgM class were detected by the newly developed ELISA in 9 of 15 sera obtained from dogs with symptoms of acute distemper. In leucocytes of 5 of the 15 dogs (all of which were also IgM positive) CDV RNA was detected by reverse transcription (RT)-PCR. The recombinant capture-sandwich ELISA detecting N-specific antibodies of the IgG class provided superior sensitivity and specificity and thus represents a rapid and cost-effective alternative to classical CDV V-NA. By detection of specific IgM antibodies, the ELISA will be complementary to RT-PCR and V-NA in the diagnosis of acute distemper infections.
PMCID: PMC88648  PMID: 10074525
10.  Detection of Canine Distemper Virus Nucleoprotein RNA by Reverse Transcription-PCR Using Serum, Whole Blood, and Cerebrospinal Fluid from Dogs with Distemper 
Journal of Clinical Microbiology  1999;37(11):3634-3643.
Reverse transcription-PCR (RT-PCR) was used to detect canine distemper virus (CDV) nucleoprotein (NP) RNA in serum, whole blood, and cerebrospinal fluid (CSF) samples from 38 dogs with clinically suspected distemper. Results were correlated to clinical findings, anti-CDV neutralizing antibody titers, postmortem findings, and demonstration of CDV NP antigen by immunohistochemistry. The specificity of the RT-PCR was ensured by amplification of RNA from various laboratory CDV strains, restriction enzyme digestion, and Southern blot hybridization. In 29 of 38 dogs, CDV infection was confirmed by postmortem examination and immunohistochemistry. The animals displayed the catarrhal, systemic, and nervous forms of distemper. Seventeen samples (serum, whole blood, or CSF) from dogs with distemper were tested with three sets of primers targeted to different regions of the NP gene of the CDV Onderstepoort strain. Expected amplicons were observed in 82, 53, and 41% of the 17 samples, depending upon the primer pair used. With the most sensitive primer pair (primer pair I), CDV NP RNA was detected in 25 of 29 (86%) serum samples and 14 of 16 (88%) whole blood and CSF samples from dogs with distemper but not in body fluids from immunohistochemically negative dogs. Nucleotide sequence analysis of five RT-PCR amplicons from isolates from the field revealed few silent point mutations. These isolates exhibited greater homology to the Rockborn (97 to 99%) than to the Onderstepoort (95 to 96%) CDV strain. In summary, although the sensitivity of the RT-PCR for detection of CDV is strongly influenced by the location of the selected primers, this nucleic acid detection system represents a highly specific and sensitive method for the antemortem diagnosis of distemper in dogs, regardless of the form of distemper, humoral immune response, and viral antigen distribution.
PMCID: PMC85712  PMID: 10523566
16.  Molecular and serological surveillance of canine enteric viruses in stray dogs from Vila do Maio, Cape Verde 
Background
Infections caused by canine parvovirus, canine distemper virus and canine coronavirus are an important cause of mortality and morbidity in dogs worldwide. Prior to this study, no information was available concerning the incidence and prevalence of these viruses in Cape Verde archipelago.
Results
To provide information regarding the health status of the canine population in Vila do Maio, Maio Island, Cape Verde, 53 rectal swabs were collected from 53 stray dogs during 2010 and 93 rectal swabs and 88 blood samples were collected from 125 stray dogs in 2011. All rectal swabs (2010 n = 53; 2011 n = 93) were analysed for the presence of canine parvovirus, canine distemper virus and canine coronavirus nucleic acids by quantitative PCR methods. Specific antibodies against canine distemper virus and canine parvovirus were also assessed (2011 n = 88).
From the 2010 sampling, 43.3% (23/53) were positive for canine parvovirus DNA, 11.3% (6/53) for canine distemper virus RNA and 1.9% (1/53) for canine coronavirus RNA. In 2011, the prevalence values for canine parvovirus and canine coronavirus were quite similar to those from the previous year, respectively 44.1% (41/93), and 1.1% (1/93), but canine distemper virus was not detected in any of the samples analysed (0%, 0/93). Antibodies against canine parvovirus were detected in 71.6% (63/88) blood samples and the seroprevalence found for canine distemper virus was 51.1% (45/88).
Conclusions
This study discloses the data obtained in a molecular and serological epidemiological surveillance carried out in urban populations of stray and domestic animals. Virus transmission and spreading occurs easily in large dog populations leading to high mortality rates particularly in unvaccinated susceptible animals. In addition, these animals can act as disease reservoirs for wild animal populations by occasional contact. Identification of susceptible wildlife of Maio Island is of upmost importance to evaluate the risk of pathogen spill over from domestic to wild animals in Cape Verde and to evaluate the associated threat to the wild susceptible species.
doi:10.1186/1746-6148-10-91
PMCID: PMC4005843  PMID: 24755118
Canine coronavirus; Canine distemper virus; Canine parvovirus; Cape verde; Molecular surveillance
17.  Improvement Control System of Rabies in Ukraine 
Objective
The purpose of the research was to find out the reasons of rabies antropurgisation in Ukraine.
Introduction
In Ukraine in spite of considerable financial expenses on oral immunization of foxes and parenteral immunization of dogs and cats, it is not succeeded to reach considerable results in the fight with rabies. Unfortunately there was a negative tendency to increasing a part of dogs and cats in the structure of rabies disease which are the main source of rabies in people.
Methods
Analysis of 228 anamnesis data of rabies infected dogs during 2008–2012. Research of 234 samples of the blood serum from dogs on existence of antibodies to the rabies virus by the ELISA method.
Results
Analysis of animal morbidity on rabies in Ukraine in period of 2006–2011 found out the changes of structure of morbidity in animal species that means decreasing a part of wild animals (from 49,0 % in 2006 to 38,7 % in 2011) and increasing a part of dogs (from 18,3 % in 2006 to 23,2 % in 2011) and cats (from 19,8 % in 2006 to 25,0 % in 2011) in the general amount of animals which perished from rabies (Fig. 1).
A lot of Ukrainian scientists and doctors of veterinary medicine consider that the main reason of spreading the rabies is a great number of homeless animals which factually are the reservoirs of infection in towns and villages.
However, in our opinion spreading of rabies shows the insufficient level of measures of control of rabies among home animals. It was confirmed with conducted analysis that only 26 (12,9 %) dogs were stray, others 202 (87,1 %) had owners, but didn’t get necessary protective rabies vaccination.
According to Ukrainian instruction “Preventive measures against rabies of animals”, all the dogs must be vaccinated against rabies, but it actually appears it is quite not so.
At research of 234 samples of the blood serum of dogs on existence of antibodies to the rabies virus it was determined that the level of population immunity in dogs is 36,6 % in Ukrainian towns, but the protective level of antibodies was found in 9,1 % of village dogs. We consider that the main reason of this is imperfection of the Ukrainian legislation in the questions of responsibility of proprietors of animals.
For breaking of epizootic chain it is necessary the percent of vaccinated animals (in this case dogs) to be on a high level – more than 50%. Nowadays for reaching epidemic welfare in rabies it is important within the framework of the registered epidemic incidents dogs to be brought over to the ecocycles of infection sporadically without taking part in circulation of exciter, stay their biological deadlock and have low epidemic potential.
Conclusions
The conducted analysis expressly demonstrates that at present problems the eradication of rabies in Ukraine is impossible considering the low level of dogs’ protection from rabies. Obtained results were sent to the State committee of veterinary medicine of Ukraine and will be the argument for enhancement of control after conducting rabies vaccination of dogs.
Rabies cases in the years 2006–2011.
PMCID: PMC3692804
population rabies immunity; rabies vaccination; stray dogs
18.  Addressing medical coding and billing part II: a strategy for achieving compliance. A risk management approach for reducing coding and billing errors. 
Medical practice today, more than ever before, places greater demands on physicians to see more patients, provide more complex medical services and adhere to stricter regulatory rules, leaving little time for coding and billing. Yet, the need to adequately document medical records, appropriately apply billing codes and accurately charge insurers for medical services is essential to the medical practice's financial condition. Many physicians rely on office staff and billing companies to process their medical bills without ever reviewing the bills before they are submitted for payment. Some physicians may not be receiving the payment they deserve when they do not sufficiently oversee the medical practice's coding and billing patterns. This article emphasizes the importance of monitoring and auditing medical record documentation and coding application as a strategy for achieving compliance and reducing billing errors. When medical bills are submitted with missing and incorrect information, they may result in unpaid claims and loss of revenue to physicians. Addressing Medical Audits, Part I--A Strategy for Achieving Compliance--CMS, JCAHO, NCQA, published January 2002 in the Journal of the National Medical Association, stressed the importance of preparing the medical practice for audits. The article highlighted steps the medical practice can take to prepare for audits and presented examples of guidelines used by regulatory agencies to conduct both medical and financial audits. The Medicare Integrity Program was cited as an example of guidelines used by regulators to identify coding errors during an audit and deny payment to providers when improper billing occurs. For each denied claim, payments owed to the medical practice are are also denied. Health care is, no doubt, a costly endeavor for health care providers, consumers and insurers. The potential risk to physicians for improper billing may include loss of revenue, fraud investigations, financial sanction, disciplinary action and exclusion from participation in government programs. Part II of this article recommends an approach for assessing potential risk, preventing improper billing, and improving financial management of the medical practice.
Images
PMCID: PMC2594405  PMID: 12078924
19.  Effect of owner-controlled acaricidal treatment on tick infestation and immune response to tick-borne pathogens in naturally infested dogs from Eastern Austria 
Parasites & Vectors  2013;6:62.
Background
Tick-borne infections resulting from regular tick infestation in dogs are a common veterinary health problem all over the world. The application of repellent and acaricidal agents to prevent transmission of pathogens is a major protection strategy and has been proven to be highly effective in several trials under laboratory and natural conditions in dogs. Despite such promising results, many dog owners still report tick infestation in their dogs although acaricidal agents are used. Information about the current infection status and changes of the infection status regarding tick-borne diseases (TBD) in dogs treated by the owner’s controlled acaricide application is lacking.
Methods
In this study 30 dogs were each treated with permethrin, fipronil + S-methoprene, or served as untreated controls. Application of the acaricide was performed by the owner who decided when and how often to use the spot on preparation. Over a period of 11 months, dogs were clinically examined and sampled for antibody responses against Babesia canis, Anaplasma phagocytophilum, Borrelia burgdorferi s. l., and TBE virus before the study started, 6 months later and at the end of the investigation period.
Results
The permethrin acaricide was applied on average 3.40 times within the examination period, whereas the fipronil + S-methoprene medication was applied 3.03 times. Approximately 2/3 of all dogs, independent of the group, had a positive immune response to one or more pathogens. Three dogs developed clinical symptoms of canine babesiosis, all other dogs remained healthy. Individual number of ticks per dog or number of infections per dog did not correlate with the application rate, and the number of ticks per dog did not influence the number of infections per dog. As owners did not apply the acaricides regularly no influence on the number of infections could be documented although the number of ticks was clearly reduced by the application of the spot-on drugs.
Conclusions
Clinical disease in dogs exposed to tick-borne pathogens is rare, although a humoral immune response reflecting infection is common. More educational training for dog owners is necessary to make the application of acaricides effective regarding the prevention of tick-borne diseases.
doi:10.1186/1756-3305-6-62
PMCID: PMC3623908  PMID: 23497548
Tick infestation; Tick-borne pathogens; Immune response; Acaricides; Efficacy
20.  Patient identified needs for chronic obstructive pulmonary disease versus billed services for care received 
The American Lung Association of Minnesota (ALAMN) was granted access to a 2004 administrative claims data from an upper mid-Western, independent practice association model health plan. Claims information, including demographics, prevalence, medication and oxygen therapy, and health care utilization, was extracted for 7,782 patients with COPD who were 40 years of age and older. In addition, ALAMN conducted a survey of 1,911 patients from Minnesota diagnosed with COPD. The survey queried the patients about demographics, treatment, medications, limitations, wants, and needs. This article compares and contrasts the information gained through the health plan administrative claims database with the findings from the COPD patient survey in areas of age, gender, types of provider primarily responsible for COPD care, spirometry use, medication therapy, pulmonary rehabilitation, oxygen therapy, and health care utilization. Primary care practitioners provided a majority of the COPD-related care. The claims evidence of spirometry use was 16%–62% of COPD patients had claims evidence of COPD-related medications. 25% of patients reported, and 23% of patients had claims evidence of, a hospitalization during the observation year. 16% of patients reported using pulmonary rehabilitation programs. The results indicate there is an opportunity to improve COPD diagnosis and management.
PMCID: PMC2629976  PMID: 18990969
chronic obstructive pulmonary disease; oxygen therapy; medication therapy; spirometry; chronic care; assessment
21.  Canine distemper virus induces apoptosis in cervical tumor derived cell lines 
Virology Journal  2011;8:334.
Apoptosis can be induced or inhibited by viral proteins, it can form part of the host defense against virus infection, or it can be a mechanism for viral spread to neighboring cells. Canine distemper virus (CDV) induces apoptotic cells in lymphoid tissues and in the cerebellum of dogs naturally infected. CDV also produces a cytopathologic effect, leading to apoptosis in Vero cells in tissue culture. We tested canine distemper virus, a member of the Paramyxoviridae family, for the ability to trigger apoptosis in HeLa cells, derived from cervical cancer cells resistant to apoptosis. To study the effect of CDV infection in HeLa cells, we examined apoptotic markers 24 h post infection (pi), by flow cytometry assay for DNA fragmentation, real-time PCR assay for caspase-3 and caspase-8 mRNA expression, and by caspase-3 and -8 immunocytochemistry. Flow cytometry showed that DNA fragmentation was induced in HeLa cells infected by CDV, and immunocytochemistry revealed a significant increase in the levels of the cleaved active form of caspase-3 protein, but did not show any difference in expression of caspase-8, indicating an intrinsic apoptotic pathway. Confirming this observation, expression of caspase-3 mRNA was higher in CDV infected HeLa cells than control cells; however, there was no statistically significant change in caspase-8 mRNA expression profile. Our data suggest that canine distemper virus induced apoptosis in HeLa cells, triggering apoptosis by the intrinsic pathway, with no participation of the initiator caspase -8 from the extrinsic pathway. In conclusion, the cellular stress caused by CDV infection of HeLa cells, leading to apoptosis, can be used as a tool in future research for cervical cancer treatment and control.
doi:10.1186/1743-422X-8-334
PMCID: PMC3141686  PMID: 21718481
Apoptosis; Canine distemper virus; Caspase; Cervical tumor; HeLa cell; HPV
22.  Review on Dog Rabies Vaccination Coverage in Africa: A Question of Dog Accessibility or Cost Recovery? 
PLoS Neglected Tropical Diseases  2015;9(2):e0003447.
Background
Rabies still poses a significant human health problem throughout most of Africa, where the majority of the human cases results from dog bites. Mass dog vaccination is considered to be the most effective method to prevent rabies in humans. Our objective was to systematically review research articles on dog rabies parenteral vaccination coverage in Africa in relation to dog accessibility and vaccination cost recovery arrangement (i.e.free of charge or owner charged).
Methodology/Principal Findings
A systematic literature search was made in the databases of CAB abstracts (EBSCOhost and OvidSP), Scopus, Web of Science, PubMed, Medline (EBSCOhost and OvidSP) and AJOL (African Journal Online) for peer reviewed articles on 1) rabies control, 2) dog rabies vaccination coverage and 3) dog demography in Africa. Identified articles were subsequently screened and selected using predefined selection criteria like year of publication (viz. ≥ 1990), type of study (cross sectional), objective(s) of the study (i.e. vaccination coverage rates, dog demographics and financial arrangements of vaccination costs), language of publication (English) and geographical focus (Africa). The selection process resulted in sixteen peer reviewed articles which were used to review dog demography and dog ownership status, and dog rabies vaccination coverage throughout Africa. The main review findings indicate that 1) the majority (up to 98.1%) of dogs in African countries are owned (and as such accessible), 2) puppies younger than 3 months of age constitute a considerable proportion (up to 30%) of the dog population and 3) male dogs are dominating in numbers (up to 3.6 times the female dog population). Dog rabies parenteral vaccination coverage was compared between “free of charge” and “owner charged” vaccination schemes by the technique of Meta-analysis. Results indicate that the rabies vaccination coverage following a free of charge vaccination scheme (68%) is closer to the World Health Organization recommended coverage rate (70%) than the achieved coverage rate in owner-charged dog rabies vaccination schemes (18%).
Conclusions/Significance
Most dogs in Africa are owned and accessible for parenteral vaccination against rabies if the campaign is performed “free of charge”.
Author Summary
Rabies is one of the most fatal diseases in both humans and animals. A bite by a rabid dog is the main cause of human rabies in Africa. Parenteral mass dog vaccination is the most cost-effective tool to prevent rabies in humans. Our main objective was to review research articles on the parenteral dog rabies vaccination coverage in Africa. We aimed to review published research articles on percentage of dogs owned and percentage of dogs vaccinated against rabies, and on the relation between vaccination coverage and cost recovery. We followed the standard procedures of a systematic literature review resulting in a final review of 16 scientific articles. Our review results indicate that only a small percentage of African dogs is ownerless. Puppies younger than 3 months of age constitute a considerable proportion of the African dog population. There are considerably more male dogs than female dogs present within the dog population. The dog rabies parenteral vaccination coverage following a “free of charge” vaccination scheme (68%) is closer to World Health Organization recommended threshold coverage rate (70%) compared to the coverage rate achieved in “owner-charged” dog rabies vaccination schemes (18%). In conclusion, most dogs in Africa are owned and accessible for vaccination once the necessary financial arrangements have been made.
doi:10.1371/journal.pntd.0003447
PMCID: PMC4315526  PMID: 25646774
23.  Contact with Domestic Dogs Increases Pathogen Exposure in Endangered African Wild Dogs (Lycaon pictus) 
PLoS ONE  2012;7(1):e30099.
Background
Infectious diseases have contributed to the decline and local extinction of several wildlife species, including African wild dogs (Lycaon pictus). Mitigating such disease threats is challenging, partly because uncertainty about disease dynamics makes it difficult to identify the best management approaches. Serious impacts on susceptible populations most frequently occur when generalist pathogens are maintained within populations of abundant (often domestic) “reservoir” hosts, and spill over into less abundant host species. If this is the case, disease control directed at the reservoir host might be most appropriate. However, pathogen transmission within threatened host populations may also be important, and may not be controllable by managing another host species.
Methodology/Principal Findings
We investigated interspecific and intraspecific transmission routes, by comparing African wild dogs' exposure to six canine pathogens with behavioural measures of their opportunities for contact with domestic dogs and with other wild dogs. Domestic dog contact was associated with exposure to canine parvovirus, Ehrlichia canis, Neospora caninum and perhaps rabies virus, but not with exposure to canine distemper virus or canine coronavirus. Contact with other wild dogs appeared not to increase the risk of exposure to any of the pathogens.
Conclusions/Significance
These findings, combined with other data, suggest that management directed at domestic dogs might help to protect wild dog populations from rabies virus, but not from canine distemper virus. However, further analyses are needed to determine the management approaches – including no intervention – which are most appropriate for each pathogen.
doi:10.1371/journal.pone.0030099
PMCID: PMC3253127  PMID: 22238695
24.  Design of different strategies of multivalent DNA-based vaccination against rabies and canine distemper in mice and dogs 
Virology Journal  2012;9:319.
Background
During the vaccination campaigns, puppies younger than 3 months old are not targeted and remain unvaccinated for at least the first year of their lives. Almost half of the reported rabid dogs are 6 months or younger. Hence, we should recommend the vaccination against rabies of young puppies. Unfortunately, owing to the exposure of puppies to infections with either canine parvovirus (CPV) or distemper virus (CDV) after the intervention of the vaccinators, owners are reluctant to vaccinate puppies against rabies. Therefore, it is necessary to include the CPV and CDV valences in the vaccine against rabies. Multivalent DNA-based vaccination in dogs, including rabies and distemper valences, could help in raising vaccine coverage.
Methods
We have designed monovalent and multivalent DNA-based vaccine candidates for in vitro and in vivo assays. These plasmids encode to the rabies virus glycoprotein and/or the canine distemper virus hemagglutinin. The first strategy of multivalent DNA-based vaccination is by mixing plasmids encoding to a single antigen each. The second is by simply fusing the genes of the antigens together. The third is by adding the foot and mouth disease virus (FMDV) 2A oligopeptide gene into the antigen genes. The last strategy is by the design and use of a bicistronic plasmid with an “Internal Ribosome Entry Site” (IRES) domain.
Results
The monovalent construct against canine distemper was efficiently validated by inducing higher humoral immune responses compared to cell-culture-derived vaccine both in mice and dogs. All multivalent plasmids efficiently expressed both valences after in vitro transfection of BHK-21 cells. In BALB/c mice, the bicistronic IRES-dependant construct was the most efficient inducer of virus-neutralizing antibodies against both valences. It was able to induce better humoral immune responses compared to the administration of either cell-culture-derived vaccines or monovalent plasmids. The FMDV 2A was also efficient in the design of multivalent plasmids.
Conclusions
In a single shot, the design of efficient multivalent plasmids will be very beneficial for DNA-based vaccination against numerous diseases.
doi:10.1186/1743-422X-9-319
PMCID: PMC3547725  PMID: 23270301
Rabies; CDV; DNA Vaccine; Multivalent; Public health; Zoonose
25.  Sand Fly Salivary Proteins Induce Strong Cellular Immunity in a Natural Reservoir of Visceral Leishmaniasis with Adverse Consequences for Leishmania 
PLoS Pathogens  2009;5(5):e1000441.
Immunity to a sand fly salivary protein protects against visceral leishmaniasis (VL) in hamsters. This protection was associated with the development of cellular immunity in the form of a delayed-type hypersensitivity response and the presence of IFN-γ at the site of sand fly bites. To date, there are no data available regarding the cellular immune response to sand fly saliva in dogs, the main reservoirs of VL in Latin America, and its role in protection from this fatal disease. Two of 35 salivary proteins from the vector sand fly Lutzomyia longipalpis, identified using a novel approach termed reverse antigen screening, elicited strong cellular immunity in dogs. Immunization with either molecule induced high IgG2 antibody levels and significant IFN-γ production following in vitro stimulation of PBMC with salivary gland homogenate (SGH). Upon challenge with uninfected or infected flies, immunized dogs developed a cellular response at the bite site characterized by lymphocytic infiltration and IFN-γ and IL-12 expression. Additionally, SGH-stimulated lymphocytes from immunized dogs efficiently killed Leishmania infantum chagasi within autologous macrophages. Certain sand fly salivary proteins are potent immunogens obligatorily co-deposited with Leishmania parasites during transmission. Their inclusion in an anti-Leishmania vaccine would exploit anti-saliva immunity following an infective sand fly bite and set the stage for a protective anti-Leishmania immune response.
Author Summary
Leishmaniasis is a neglected infectious disease with a global distribution encompassing 88 countries, 350 million people at risk, and an annual incidence of 2 million cases. Leishmaniasis is a vector-borne disease transmitted by sand fly bites where parasites are co-deposited with saliva into the wound. Our group has demonstrated that distinct molecules in the saliva of various sand fly species drive an immune response that protects experimental rodent models from self-healing cutaneous and fatal visceral leishmaniasis. Here we show for the first time that dogs, natural reservoirs of visceral leishmaniasis, develop a strong immune response to two salivary proteins from the natural vector sand fly. Blood from immunized dogs contained immune cells that produced molecules (IFN-γ) typically associated with protection from Leishmania parasites. This response efficiently recruited appropriate immune cells to the site of sand fly bites in the skin and had an adverse effect on Leishmania parasites in an experimental assay. These findings suggest that inclusion of these salivary molecules in anti-Leishmania canine vaccines would enhance their efficiency in protecting dogs from visceral leishmaniasis. A successful anti-Leishmania canine vaccine would not only protect dogs from a fatal disease but could have a considerable effect on reducing human infections.
doi:10.1371/journal.ppat.1000441
PMCID: PMC2677456  PMID: 19461875

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