A COMPARISON OF THE OBJECTIVE STRUCTURED CLINICAL EXAMINATION AND THE TRADITIONAL ORAL CLINICAL EXAMINATION IN A NIGERIAN UNIVERSITY.

BACKGROUND: Assessment of clinical skills is essential in medical education. Ideally marks should be based on the student's competence alone. The limitations of the traditional long case examinations such as the patient and examiner variability are well known. The objective structured clinical examination (OSCE) was designed to overcome these limitations. Studies comparing the OSCE and the traditional long case examination in the same group of students are very sparse. AIM: To compare the objective structured clinical examination (OSCE) and the traditional long case examination by determining their correlations with other forms of assessment in undergraduate surgery. SETTING: This study was carried out at the College of Medicine, University of Lagos, Nigeria. METHODOLOGY: The results of 612 undergraduate students in our medical school of the University of Lagos, Nigeria over a period of 4 years (2012-2015) were analysed. The scores in the long case examination , objective structured clinical examination (OSCE) , multiple choice questions and Essays were analysed and compared using the Pearson's Correlation co-efficient. SPSS version 17 software was used and a P-value < 0.01 was regarded as statistically significant. RESULTS: Overall, there was a statistical significant positive correlation among most forms of assessment. The OSCE and the long case examination had a correlation of 0.374. Compared with the long case examination, the OSCE had a higher correlation with all other forms of assessment. The total clinical score (the sum of all long case examination and OSCE) however performed better than the OSCE or the long case examination alone as it had the highest correlation with all other forms of assessment. CONCLUSION: The OSCE has been shown to be better than the long case examination as an indicator of the student overall knowledge of surgery as it had a superior correlation with other forms of assessments. The total clinical score was however the best indicator of the student overall knowledge in Surgery as it had the best correlation with other forms of assessment. We recommend and encourage institutions that presently combine the OSCE and the long case examination to carry out similar analysis such as ours to determine the desirability of combining LCE and OSCE rather than outright replacement of LCE with OSCE.

Control of colonization by virulent Salmonella typhimurium by oral immunization of chickens with avirulent delta cya delta crp S. typhimurium.

Oral immunization with a delta cya delta crp Salmonella typhimurium strain has been shown to preclude colonization by wild-type, virulent S. typhimurium and induces humoral and cellular immune response in chickens. Intestinal tract colonization by the virulent challenge strain was used to determine the level of protection conferred by immunization with the delta cya delta crp mutant. The associated humoral and cellular immune responses were measured by ELISA and delayed-type hypersensitivity (DTH) tests, respectively. The levels of colonization by both Salmonella strains were determined by enumeration of viable cells in the intestinal tract. A reduction in faecal excretion of the wild-type strain was observed with a single oral immunization with the delta cya delta crp mutant, but caecal colonization was not affected. However, double oral immunization with the delta cya delta crp mutant precludes caecal colonization by the virulent strain. IgM, IgA and IgG were detected against sonicated Salmonella whole-cell antigens. Outer membrane and flagella proteins induced DTH responses, whereas lipopolysaccharide failed to do so. The effectiveness of the delta cya delta crp strain in reducing caecal colonization by the highly virulent challenge strain in chickens demonstrates that oral vaccination with the delta cya delta crp S. typhimurium should aid in eliminating Salmonella carriers in chickens. The elimination of these carriers on the poultry farm should help to control Salmonella contamination of poultry products, therapy improving public health.

Live oral avirulent Salmonella vaccines.

Infection of animals and humans with Salmonella is a consequence of oral consumption of food or fluids contaminated with Salmonella. Once in the intestine, Salmonella usually attach to, invade, and proliferate in enterocytes or the cells of the gut associated lymphoid tissue (GALT). The latter route of infection can lead to disease or to an asymptomatic carrier state or stimulate the induction of mucosal, systemic and cellular immune responses. Infection of animals with virulent invasive Salmonella can result in suppression of the immune responses which in turn can facilitate the establishment of a carrier state. It is possible to attenuate Salmonella by introducing mutations that (i) confer auxotrophy, (ii) interfere with sugar metabolism and LPS biosynthesis or (iii) affect some global means of regulating genes needed for the full display of virulence. Oral immunization of animals such as mice and chickens with avirulent Salmonella strains usually is not associated with suppression but rather with stimulation of mucosal, systemic and cellular immune responses. Vaccination by injection of killed vaccines or bacterins does not lead to the induction of either mucosal or cellular immune responses, and humoral immunity may be relatively short lived. Thus, killed vaccines are inferior to orally administered live avirulent Salmonella vaccines which induce a long-lasting protective immunity. In this manuscript we discuss desirable attributes of a safe, efficacious live attenuated Salmonella vaccine, describe attenuated Salmonella mutants so far isolated and their properties and present information on the evaluation of a live attenuated Salmonella oral vaccine for poultry.(ABSTRACT TRUNCATED AT 250 WORDS)

Nonrecombinant and recombinant avirulent Salmonella vaccines for poultry.

We have constructed and evaluated a live avirulent Salmonella typhimurium vaccine strain, attenuated by deletion (delta) mutations in genes for adenylate cyclase (cya) and the cAMP receptor protein (crp). Immunization of chicks can preclude Salmonella colonization and invasion of challenged vaccinated chickens when compared with the non vaccinated control. Immunization induces significant cross-protective immunity against various Salmonella serotypes and protects laying hens from transmission of Salmonella in or on eggs following challenge with S. enteritidis or S. typhimurium. Immunization of chicks destined to be breeders and then with a booster immunization at 16-18 weeks of age leads to maternal transfer of immunity to chicks which then can be immunized either orally or by coarse spray to display an enhanced immunity to prevent infection of visceral organs by and shedding of Salmonella. The attenuated S. typhimurium vaccine can be genetically manipulated to express foreign antigens specified by cloned genes from other pathogens. Immunization with such recombinant vaccines not only induces immunity to Salmonella but to infection by the pathogen that supplied the genes specifying the protective antigens expressed by the recombinant vaccine.

Efficacy of a live avirulent Salmonella typhimurium vaccine in preventing colonization and invasion of laying hens by Salmonella typhimurium and Salmonella enteritidis.

An avirulent live delta cya delta crp Salmonella typhimurium strain chi 3985 that precludes colonization and invasion of chickens by homologous and heterologous Salmonella serotypes was evaluated for its long-term protection efficacy. Chickens vaccinated orally at 2 and 4 wk of age were assessed for protection against oral challenge with wild-type S. typhimurium and Salmonella enteritidis strains at 3, 6, 9, and 12 mo of age. A comparison of Salmonella isolation from vaccinated and nonvaccinated layers after challenge with S. typhimurium or S. enteritidis showed that delta cya delta crp S. typhimurium chi 3985 induced excellent protection against intestinal, visceral, reproductive tract, and egg colonization, invasion, and/or contamination by Salmonella. The duration of protection lasted for 11 mo after vaccination, at which time the experiment was terminated. S. enteritidis and S. typhimurium were isolated from the yolk, albumen, and shells of eggs laid by nonvaccinated chickens challenged with Salmonella. S. typhimurium caused pathological lesions in nonvaccinated chickens, whereas vaccinated and nonvaccinated chickens challenged with S. enteritidis showed no pathological lesion in the visceral and reproductive organs. Vaccination with chi 3985 prevented transmission of S. typhimurium or S. enteritidis into eggs laid by vaccinated layers with no effect on egg production. To our knowledge, this is the first publication confirming that vaccination with live avirulent Salmonella can induce long-term protection against Salmonella infection in layers.

Effect of infective dose on humoral immune responses and colonization in chickens experimentally infected with Salmonella typhimurium.

The influence of infective dose on chicken immunogenicity was examined in 1-week-old chickens. Chickens were infected orally with various doses of chi 3761 or chi 3985. Fecal shedding, colonization of the cecum, and induction of Salmonella-specific serum immunoglobulin isotypes were analyzed over a 5-week period. The delta cya delta crp Salmonella typhimurium vaccine strain chi 3985 was used to assess the effect of vaccination dose on protection after oral vaccination of chickens at 1 day and 2 weeks of age. Wild-type S. typhimurium strain chi 3761 was used to challenge vaccinated and unvaccinated chickens at 6 weeks of age, and the recovery of Salmonella from the cecum was used as a measure of protection. Infection of 1-week-old chickens with chi 3985 was more effective in reducing fecal excretion and cecal colonization than was infection with chi 3761. Double vaccination with 10(8) or 10(7) CFU of chi 3985 at 1 day and 2 weeks of age protected vaccinated chickens against cecal colonization by the challenge strain chi 3761. Immunogenicity of Salmonella is dose and genotype-dependent.

Infection and reinfection of chickens with Salmonella typhimurium: bacteriology and immune responses.

Four-day-old chickens infected orally with a spectinomycin-resistant (Spcr) mutant of a highly invasive avian Salmonella typhimurium strain excreted salmonellae in the feces for at least 10 weeks. When these chickens were reinfected at this time with a nalidixic acid-resistant (Nalr) mutant of the same strain, they excreted this mutant in significantly smaller numbers (P less than 0.01) than did a previously uninfected control group. The Nalr mutant had a shorter survival rate in the tissues of the immunized chickens than in tissues of the control birds. The Spcr mutant stimulated strong IgG, IgA, and IgM responses in serum, small-intestinal contents, and bile. These were detected by enzyme-linked immunosorbent assay (ELISA) against antigens of crude whole bacterial cell protein sonicate, lipopolysaccharide, flagella, and outer-membrane proteins. There was some evidence of an anamnestic response with IgA in bile following reinfection with the Salmonella. The peak response of antibody-producing cells from the spleens of infected chickens, assayed by solid-phase ELISA, occurred at 3 weeks postinoculation. A strong delayed hypersensitivity reaction, detected by foot-pad swelling after inoculation with either whole-cell or outer-membrane proteins, was observed between 2 and 5 weeks after infection with the Spcr mutant. The data indicate that outer-membrane proteins are major immunogens for both humoral and cell-mediated arms of the immune system.

Antibody response to experimental Salmonella typhimurium infection in chickens measured by ELISA.

An indirect ELISA has been developed to detect Salmonella typhimurium antibodies in chicken sera, using whole bacterial cell protein, flagellar protein or lipopolysaccharide as antigens. In experimental infections high concentrations of S typhimurium-specific IgG persisted after the faecal excretion of S typhimurium had ceased, whereas the specific IgM response was transitory. Some uninfected chickens placed in contact with experimentally infected birds developed high IgG titres in the absence of detectable faecal excretion. Other S typhimurium strains, which varied in their invasive abilities, also induced high titres of IgG. The ELISA allowed chickens infected experimentally with S typhimurium to be differentiated from chickens infected with 10 other serotypes, including S enteritidis. The use of whole blood in place of serum in the ELISA reduced the titres slightly. The storage of serum dried on to filter paper strips for four weeks produced little change in ELISA antibody titre, and the treatment of such strips with phenol or chloroform vapour had little or no effect on the antibody titre.

A survey of the bacteriological status of fresh and baked pork sausages produced in Ibadan, Nigeria.

A survey was conducted on the bacteriological status of fresh and baked pork sausages produced in Ibadan Nigeria, and to assess the associated public health hazards, using Salmonella spp, coagulase-positive Staphylococci and enteropathogenic Escherichia coli as reference organisms. Salmonellae were present in 42% of the fresh pork sausage samples and in 56% of the baked samples. Coagulase-positive Staphylococcus aureus was identified in 16% of the fresh sausage samples and in 26% of baked samples, while Escherichia coli isolation rates were 48% and 19% in fresh and baked sausages respectively. To protect public health, the importance of improved systems of lairaging, attention to methods of transport, reduction of stress in the animals, improved methods of slaughter; particularly in evisceration as well as hygiene of utensils and meat handlers are emphasised.

Virulent Salmonella typhimurium-induced lymphocyte depletion and immunosuppression in chickens.

The effect of experimental Salmonella infection on chicken lymphoid organs, immune responses, and fecal shedding of salmonellae were assessed following oral inoculation of 1-day-old chicks or intra-air-sac infection of 4-week-old chickens with virulent S. typhimurium wild-type chi 3761 or avirulent S. typhimurium delta cya delta crp vaccine strain chi 3985. Some 4-week-old chickens infected intra-air-sac with chi 3761 or chi 3985 were challenged with Bordetella avium to determine the effect of Salmonella infection on secondary infection by B. avium. S. typhimurium chi 3761 caused lymphocyte depletion, atrophy of lymphoid organs, and immunosuppression 2 days after infection in 1-day-old chicks and 4-week-old chickens. The observed lymphocyte depletion or atrophy of lymphoid organs was transient and dose dependent. Lymphocyte depletion and immunosuppression were associated with prolonged fecal shedding of S. typhimurium chi 3761. No lymphocyte depletion, immunosuppression, or prolonged Salmonella shedding was observed in groups of chickens infected orally or intra-air-sac with chi 3985. Infection of chickens with salmonellae before challenge with B. avium did not suppress the specific antibody response to B. avium. However, B. avium isolation was higher in visceral organs of chickens infected with chi 3761 and challenged with B. avium than in chickens infected with B. avium only. Infection of chickens with chi 3985 reduced B. avium colonization. We report a new factor in Salmonella pathogenesis and reveal a phenomenon which may play a critical role in the development of Salmonella carrier status in chickens. We also showed that 10(8) CFU of chi 3985, which is our established oral vaccination dose for chickens, did not cause immunosuppression or enhance the development of Salmonella carrier status in chickens.

Development and evaluation of an experimental vaccination program using a live avirulent Salmonella typhimurium strain to protect immunized chickens against challenge with homologous and heterologous Salmonella serotypes.

A stable live avirulent, genetically modified delta cya delta crp Salmonella typhimurium vaccine strain, chi 3985, was used in several vaccination strategies to evaluate its use in the control of Salmonella infection in chickens. Oral vaccination of chickens at 1 and at 14 days of age with 10(8) CFU of chi 3985 protected against invasion of spleen, ovary, and bursa of Fabricius and colonization of the ileum and cecum in chickens challenged with 10(6) CFU of virulent homologous Salmonella strains from group B. Chickens challenged with heterologous Salmonella strains from groups C, D, and E were protected against visceral invasion of spleen and ovary, while invasion of the bursa of Fabricius and colonization of ileum and cecum was reduced in vaccinated chickens. Oral vaccination at 2 and at 4 weeks of age induced an excellent protection against challenge with virulent group B Salmonella serotypes and very good protection against challenge with group D or E Salmonella serotypes, while protection against challenge with group C Salmonella serotypes was marginal but significant. Vaccination at 2 and at 4 weeks of age also protected vaccinated chickens against challenge with 10(8) CFU of highly invasive S. typhimurium or S. enteritidis strains. The protection of chickens vaccinated with chi 3985 against challenge with homologous and heterologous Salmonella serotypes is outstanding, and the complete protection against ovarian invasion in chickens challenged with 10(8) CFU of highly invasive S. typhimurium or S. enteritidis strains suggests that vaccination of chickens with chi 3985 can complement the present hygiene- and sanitation-based Salmonella control measures. This paper reports a breakthrough in the use of live avirulent vaccine to control Salmonella carriers in chickens.

Effect of vaccination of hens with an avirulent strain of Salmonella typhimurium on immunity of progeny challenged with wild-Type Salmonella strains.

The avirulent Salmonella typhimurium chi3985 was used to vaccinate white leghorn chickens at 16 and 18 weeks of age, and the effect of maternal antibody on Salmonella colonization of progeny of vaccinated hens was assessed with S. typhimurium F98 or chi3985. Progeny of hens that had been vaccinated at 1 and 3 or 2 and 4 weeks of age with chi3985 were used to determine the effect of maternal immunity on vaccine efficacy. Vaccination of hens induced long-lasting Salmonella-specific antibodies which were transferred into eggs and were detected as immunoglobulin G (IgG) in the egg yolk. Maternal antibody was detected in the progeny of vaccinated birds as IgG and IgA in serum and intestinal fluid, respectively. The titer of maternally transmitted IgG or IgA was highest in the first week of life of the progeny and declined with age. Maternal antibodies prevented colonization of the chicks by S. typhimurium chi3985 and reduced colonization by S. typhimurium F98. Overall, chicks from vaccinated hens had significantly higher antibody responses than did the progeny of nonvaccinated hens after oral infection with Salmonella strains. Maternal antibody reduced the efficacy of vaccination of progeny with chi3985 at 1 and 3 weeks of age. But vaccination at 2 and 4 weeks of age induced excellent protection against challenge with S. typhimurium F98 or S. enteritidis 27A PT 8 in birds from vaccinated hens and in specific-pathogen-free chickens. Vaccination of chickens at 2 and 4 weeks of age has been shown to protect the birds against challenge with homologous and heterologous Salmonella serotypes. A combination of vaccination of adult animals and use of the progeny of vaccinated birds will enhance effective control of Salmonella infections in the poultry industry. This will complement the present control of Salmonella-associated food poisoning caused by Salmonella enteritidis in eggs because the avirulent S. typhimurium vaccine strain chi3985 induced excellent protection against S. enteritidis in chickens.

Public health importance of market meat exposed to refuse flies and air-borne microorganisms.

Abbatoir meats sold in the open at the Ibadan market stalls were swabbed to investigate the presence of any microorganism. Flies were caught on carcases being transported from the abbatoir in open vehicles to meat stalls and also at meat stalls and nearby refuse using fly catching nets. Major flies caught were Musca domestica; Culicoides species, Chrysomia and Fannia cannicularis. Stophylococcus aureus, Salmonella and Escherichia coli were the bacteria isolated from.

Reduction in faecal excretion of Salmonella typhimurium strain F98 in chickens vaccinated with live and killed S. typhimurium organisms.

Chickens given orally at 4 days of age a smooth spectinomycin resistant mutant (Spcr) of Salmonella typhimurium strain F98 excreted the organism in their faeces for approximately 4 weeks. Following oral administration of a nalidixic acid resistant (Nalr) mutant of the same strain 4 weeks later when the chickens had virtually cleared themselves of the first infection, these chickens excreted far fewer salmonella organisms and for a shorter time than did a previously uninfected control group of chickens which were infected at the same time with the Nalr mutant. Chickens inoculated intramuscularly at 4 days developed a similar immunity to challenge and also excreted the immunizing strain in their faeces. In contrast intramuscular inoculation or incorporation into the food of formalin-killed S. typhimurium organisms had little lasting effect on the faecal excretion of the challenge strain. Two attenuated mutants of strain F98 Nalr were produced: one was a rough strain produced by lytic bacteriophage and the other was an aro A auxotrophic mutant which had been cured of the 85 kilobase-pair virulence-associated plasmid. These mutants were avirulent for chickens, mice, calves and man and when ingested by human volunteers did not persist in the faeces. When inoculated intramuscularly into chickens they produced an early reduction in faecal excretion of the challenge strain (Spcr) which was not maintained. Oral administration of both strains produced reductions in faecal excretion of the challenge strain. This was much more noticeable with the rough strain which was itself excreted for a much longer period than the parent strain.

Indirect antigen-trap ELISAs using polyclonal antisera for detection of group B and D Salmonellas in chickens.

Indirect antigen-trap ELISAs able to detect Salmonella typhimurium and S. enteritidis are described. Their limits of sensitivity were about 10(7) organisms in nutrient broth culture. With the ELISA developed for S. typhimurium, 76 of 77 strains gave optical density (OD) readings of 1.06 to 1.40. Three other serotypes from group B in the Kaufmann-White scheme gave high values: S. saintpaul produced an OD of 1.40, S. schwarzengrund 0.93 and S. derby 0.88. Twenty other serotypes all produced OD values of 0.39 to 0.66. Four Citrobacter strains and an E. coli produced lower values. With the S. enteritidis ELISA, seven strains of this serotype produced OD values of 1.13 to 1.23 and 13 other serotypes gave OD values of 0.14 to 0.87. There was good correlation between the ELISA and bacteriological culture after examining selenite broth cultures of cloacal swabs taken from experimentally infected chickens. A few samples were positive by ELISA and negative by culture and vice versa. Similar results were obtained from cloacal swabs taken from a commercial flock known to be infected with S. typhimurium. Much closer correlation existed between the two methods for identifying artificially contaminated eggs or for spleens taken from experimentally infected birds.