In vitro assessment of pathogenicity and virulence encoding gene profiles of avian pathogenic Escherichia coli strains associated with colibacillosis in chickens.

BACKGROUND: Avian pathogenic Escherichia coli (APEC) strains have been associated with various disease conditions in avian species due to virulence attributes associated with the organism. AIMS: This study was carried out to determine the in vitro pathogenic characteristics and virulence encoding genes found in E. coli strains associated with colibacillosis in chickens. METHODS: Fifty-two stock cultures of E. coli strains isolated from chickens diagnosed of colibacillosis were tested for their ability to produce haemolysis on blood agar and take up Congo red dye. Molecular characterization was carried out by polymerase chain reaction (PCR) amplification of virulence encoding genes associated with APEC. RESULTS: Eleven (22%) and 41 (71%) were positive for haemolysis on 5% sheep red blood agar and Congo red agar, respectively. Nine virulence-associated genes were detected as follows: FimH (96%), csgA (52%), iss (48%), iut (33%), tsh (21%), cva (15%), kpsII (10%), pap (2%), and felA (2%). CONCLUSION: The APEC strains exhibited virulence properties and harbored virulence encoding genes which could be a threat to the poultry population and public health. The putative virulence genes were diverse and different in almost all isolate implying that pathogenesis was multi-factorial and the infection was multi-faceted which could be a source of concern in the detection and control of APEC infections.

Pathology and Distribution of Velogenic Viscerotropic Newcastle Disease Virus in the Reproductive System of Vaccinated and Unvaccinated Laying Hens (Gallus gallus domesticus) by Immunohistochemical Labelling.

This study investigated the pathological changes in the reproductive system of laying hens that lead to the poor egg production and quality in Newcastle disease (ND) and the distribution of the virus in the system. Two hundred and forty Isa-Brown pullets were divided randomly into vaccinated and unvaccinated groups (n = 120 each). The vaccinated group was given Hitchner B1 vaccine at 1 day of age, La Sota vaccine at 4 weeks of age and Komarov vaccine at 9 and 16 weeks of age. At the peak of egg production, the laying hens (32 weeks old) were assigned randomly into four groups (n = 60): VC, vaccinated with ND vaccines and inoculated intramuscularly with velogenic viscerotropic ND virus (vvNDV); VU, vaccinated unchallenged; UC, unvaccinated challenged; and UU, unvaccinated unchallenged. UC hens showed depression, diarrhoea and later torticollis. Mortality in UC hens was 90%. VC hens showed mild anorexia. The body weights of the UC hens were significantly (P <0.05) lower than those of UU hens. VC and UC hens showed a significant (P <0.05) drop in egg production. Only UC hens produced abnormal eggs and initially had swollen, oedematous, hyperaemic oviducts followed by atrophy and shortening of the reproductive tract with atresia of the ovarian follicles. The histopathological changes were of necrosis of the epithelium and secretory glands. VC hens showed mild inflammatory changes in the oviduct. Immunohistochemical labelling showed extensive presence of the virus in the ovary, infundibulum, magnum, isthmus, uterus and vagina of UC hens and in the ovary of VC hens. These changes will be the cause of serious egg production problems, especially in vaccinated layers in countries where vvNDV is enzootic.

Preliminary evaluation of the immunoenhancement potential of Newcastle disease vaccine formulated as a cationic liposome.

This study evaluates the enhancement of immune response of birds to Newcastle disease (ND) vaccine encapsulated in 1,2-dioleoyl-3-trimethylammonium propane (DOTAP)-based liposomes. The vesicles of the liposomal ND vaccine were physically characterized for shape, particle size and zeta potential. The results of the analyses showed that vesicles of the liposomal ND vaccine were spherical and tightly packed. The mean size distribution was below 100 nm. The mean zeta potential was 24 mV. Sixty experimental birds were then divided into an unvaccinated group, a liposomal ND vaccine group and a live La Sota(®) vaccine group. Both the liposomal ND vaccine and live La Sota(®) vaccine groups were vaccinated orally at 3 and 6 weeks of age. The mean antibody titres, total and differential white blood cell count, and blood chemistry, respectively, were assessed. Ten birds from each group were challenged by oral administration of 0.2 ml virulent Herts 33 strain at 9 weeks of age. The log(2) mean antibody titre induced by the liposomal ND vaccine after secondary immunization of the birds was 9.60±0.95 while that of the live La Sota( (®) ) vaccine was 6.00±0.63. Nine of the 10 challenged birds in the unvaccinated group died while none died from the liposomal ND vaccine group or the live La Sota(®) vaccine group. After the boost vaccination, the chickens vaccinated with the liposomal ND vaccine had a higher mean antibody titre, indicating that encapsulating ND vaccine in DOTAP-based liposome induced significantly higher immunity than the live La Sota(®) vaccine.

LaSota vaccination may not protect against the lesions of velogenic Newcastle disease in chickens.

Two groups of six weeks old cockerels comprising 40 immunized and 40 non-immunized birds were inoculated intramuscularly with VGF-1, which is a local Nigerian strain of velogenic Newcastle disease virus (VNDV). Immunized birds did not show any clinical signs except significant loss (p < 0.05) in body weight on days 5 and 20 post inoculation (PI). But the non-immunized birds showed clinical signs of disease characterized by anorexia and drowsiness from day 2 PI. These were followed on day 3 PI by depression, diarrhoea, opisthotonus, weight loss (p < 0.05) and high mortalities (96.9%). Both the immunized and non-immunized groups showed severe atrophy of the bursa, spleen and thymus. Histopathological section of these lymphoid organs showed necrosis and depletion of lymphocytes. Both the gross and microscopic lesions were more severe in the non-immunized birds. Marked ballooning degeneration was observed in the bursal follicles of the non-immunized birds. This lesion has not been described earlier for any other disease and could be diagnostic for VND. Our results also showed that VND can cause marked atrophy of the lymphoid organs, which may lead to immunosupression without the characteristic signs of Newcastle disease (ND) in vaccinated chickens. This no doubt emphasizes the limitation of vaccination as a biosecurity measure in poultry industry.

The leucocytic and parasitaemic profiles and immune response of rats treated with retinyl palmitate before infection with Trypanosoma brucei.

The effects of oral administration of retinyl palmitate (30,000 IU/kg bodyweight) for 3 days to rats before infection with Trypanosoma brucei were investigated by examining the leucocytic and parasitaemic profiles, and the antibody response to sheep red blood cells. The pretreatment significantly (P<0.01) improved the leucocytic profile (especially the absolute lymphocyte count) from day 7 post-infection to the time of death. It also significantly delayed the onset of parasitaemia (i.e., lengthened the pre-patent period) and led to reduced levels of parasitaemia throughout the period of infection. Pretreatment significantly increased the antibody response to sheep red blood cells (P<0.01) throughout the infection, even after treatment with diminazene aceturate, to levels that more than compensated for the immunosuppressive effect of the T. brucei. Further studies are warranted on the possible use of retinyl palmitate in overcoming immunosuppression associated with trypanosome infections in man and animals in endemic areas.