Identification of Mycoplasma mycoides subspecies mycoides from slaughtered cattle in two transboundary states of North­eastern Nigeria.

This study aimed to perform molecular typing of Mycoplasma mycoides subsp. mycoides from slaughtered cattle in Adamawa and Taraba States, north­eastern Nigeria. A total of four hundred and eighty (480) samples of lung tissues, nasal swabs, ear swabs and pleural fluids were collected from cattle at slaughter and processed according to standard laboratory protocols. Identification and confirmation were achieved with specific PCR and PCR­RFLP. An overall M. mycoides subsp. mycoides isolation rate of 6.87% (33/480) was obtained. In Adamawa State, 12 (10.91%) isolates of M. mycoides subsp. mycoides came from both, lung tissues and pleural fluids. While in Taraba State, 5 (7.14%) and 4 (5.71%) isolates of M. mycoides subsp. mycoides came from lung tissues and pleural fluids, respectively. The samples from nasal and ear swabs from the study states were negative for M. mycoides subsp. mycoides. Thirty­three out of the 37 culture positive isolates were confirmed to be Mycoplasma mycoides subspecies mycoides with the production of a band equivalent to 574­bp. Molecular typing with restriction endonuclease Vsp1 results in the two bands of 180­bp and 380­bp. In conclusion, the study has established an isolation rate of 6.87% for M. mycoides subsp. mycoides. Measures to strengthen movement control in order to minimise the spread of this dreaded disease of cattle were recommended.

Quantitative distribution and interaction of Salmonella Zega with host cells in visceral organs of chickens infected orally, intraperitoneally and per cloaca.

Immunohistochemical study of the visceral organs of chickens experimentally infected with Salmonella Zega by three routes was carried out to compare the quantitative distribution and interaction of the organism with host cells. 100 birds comprising of 2 week-old chickens were divided into 4 groups of 25 each. Group A was inoculated orally, group B intraperitoneally, group C were administered per cloaca and D were not inoculated and served as control. All the infected birds were inoculated with 0.2 ml of 1 × 108 cfu of the bacteria. Two birds from each group were sacrificed every 24 h post infection. Samples of visceral organs were collected for immunohistochemistry. The distribution of Salmonella Zega in every organ was taken as Mean ± SD of the number of foci of immunoreactions and Compared using a 2-way ANOVA. The interaction of Salmonella Zega with host cells was determined by taking the percentage of the days post infection in which immunoreactions were detected in host cells in each route of infection. The distribution of the organism was highest in the lung of intraperitoneally infected chickens (83.95 ± 27.89) and lowest in the heart (5.21 ± 3.65) of chickens that were infected per cloaca. The highest percentage of interaction of Salmonella Zega was recorded in the epithelial (100%) and blood (100%) cells in all the routes of infection. There were variations in the distribution of Salmonella Zega in visceral organs of chickens but the level of interactions with host cells were similar even when infected through different routes.

Comparison of the clinical signs, pathological and immuohistochemical findings in visceral organs of chickens experimentally infected with Salmonella Zega through three routes.

Salmonella Zega isolated from natural outbreaks that were characterized by high mortality in poultry farms in three Southwestern States of Nigeria was used to inoculate two week-old chicks through different routes in order to determine and compare the clinical signs, pathological and immunohistochemical changes in each route of infection. The birds were divided into 4 groups of 25 each as groups A (orally inoculated), B (intraperitoneally inoculated), C (inoculated per cloaca) and D (uninoculated control). All the birds were inoculated with 0.2 ml of 1 × 108 cfu of the bacteria. Clinical signs were observed and recorded according to the route of infection, and with the days post-infection from day 0 till day 10 post-infection. Two birds from each group were sacrificed every 24 h and examined for gross lesions, which were described and scored according to the route of infection and days post-infection. Samples of visceral organs were collected for bacteriology, histopathology and immunohistochemistry. Clinical signs in chicks infected orally and intraperitoneally were weakness, anoraexia lethargy, somnolescence, yellowish diarrhoea observed from 4 days till day 10 post infections. Mild sign of weakness was observed in chickes infected per cloaca, from day 3 to 7. The gross lesions were congestion, oedema and enlargement and necrosis in visceral organs from day 4 to 10 post infection in orally and intraperitoneally infected chicks, but mild vascular changes were observed in chicks infected per cloaca, except in the caecum were lesions of necrosis and infiltration of inflammatory cells were moderate to severe. Microscopic lesions were necrosis of host cells and infiltration by lymphocytes, heterophils and macrophages in multiple organs observed from day 4 to 10 post infection in orally and intraperitoneally infected chicks. Immunoreactions were observed in all the visceral organs examined. Clinical signs, pathological and immunohistochemical findings were mild in chicks infected per cloaca, except caecal lesions. Salmonella Zega isolated from an outbreak in poultry farms in Abeokuta, Nigeria was highly pathogenic in chicken and produced similar findings in oral and intraperitoneal infections; while per cloacal infection showed a localized infection of the caecum.

Motile Salmonella serotypes causing high mortality in poultry farms in three South-Western States of Nigeria.

This study was carried out to identify the Salmonella serotypes causing high mortality in chickens in Lagos, Ogun and Oyo states, Nigeria. Chickens presented for postmortem examination during disease outbreaks that were characterised by high mortality (40 per cent to 80 per cent) in poultry farms in the study area were examined from January to December, 2013. Samples of the lungs, heart, liver, spleen, kidneys, proventriculus, intestine and caecum were collected from suspected cases of salmonellosis, for bacterial culture and identification. Salmonella isolates were confirmed using PCR and serotyped using the Kauffman-White scheme. Twenty-six day-old pullets were raised to two weeks and inoculated orally with 0.2 mL of 1×108 colony forming units of Salmonella Zega identified in the present study to determine their pathogenicity, while another 26 served as control. The Salmonella serotypes were S Zega (n=13; 35.14 per cent), Salmonella Kentucky (n=9; 24.32 per cent), Salmonella Herston (n=6; 16.22 per cent), Salmonella Nima (n=4; 10.81 per cent), Salmonella Telelkebir (n=3; 8.11 per cent), Salmonella Colindale (n=1; 2.70 per cent) and Salmonella Tshiongwe (n=1; 2.70 per cent). Clinical signs in both natural and experimental infections were acute (70 per cent) and chronic (30 per cent), and included weakness, anorexia, yellowish diarrhoea, pasted vents, somnolescence and mortality, while gross lesions showed marked pulmonary congestion and oedema, necrotic foci in the myocardium; the liver, spleen and kidneys were markedly enlarged and had subcapsular multifocal necrosis. There were catarrhal proventriculitis and enteritis, and haemorrhagic typhlitis. While most of the serotypes identified in the present study have been isolated from poultry sources from commercial farms in Nigeria, to the best of the authors' knowledge, they have not been previously reported to cause high mortality in chickens in the study area.

Antimicrobial resistance and virulence profile of enterococci isolated from poultry and cattle sources in Nigeria.

This study investigated the occurrence, antimicrobial resistance and virulence of Enterococcus from poultry and cattle farms. Three hundred and ninety samples: cloacal/rectal swabs (n = 260) and manure (n = 130] were processed for recovery of Enterococcus species. Standard bacteriological methods were used to isolate, identify and characterize Enterococcus species for antimicrobial susceptibility and expression of virulence traits. Detection of antibiotic resistance and virulence genes was carried out by polymerase chain reaction. Enterococcus was recovered from 167 (42.8%) of the 390 samples tested with a predominance of Enterococcus faecium (27.7%). Other species detected were Enterococcus gallinarum, Enterococcus faecalis, Enterococcus hirae, Enterococcus raffinosus, Enterococcus avium, Enterococcus casseliflavus, Enterococcus mundtii and Enterococcus durans. All the isolates tested were susceptible to vancomycin, but resistance to tetracycline, erythromycin, ampicillin and gentamicin was also observed among 61.0, 61.0, 45.1 and 32.7% of the isolates, respectively. Sixty (53.1%) of the isolates were multidrug resistant presenting as 24 different resistance patterns with resistance to gentamicin-erythromycin-streptomycin-tetracycline (CN-ERY-STR-TET) being the most common (n = 11) pattern. In addition to expression of virulence traits (haemolysin, gelatinase, biofilm production), antibiotic resistance (tetK, tetL, tetM, tetO and ermB) and virulence (asa1, gelE, cylA) genes were detected among the isolates. Also, in vitro transfer of resistance determinants was observed among 75% of the isolates tested. Our data revealed poultry, cattle and manure in this area are hosts to varying Enterococcus species harbouring virulence and resistance determinants that can be transferred to other organisms and also are important for causing nosocomial infection.