Isolation and identification of Mycoplasma mycoides subsp. mycoides in cattle from south-east Nigeria.

Background: Mycoplasma mycoides subsp. mycoides is the causative organism of Contagious Bovine Pleuropneumonia (CBPP). It is a trans-boundary disease and an endemic in Nigeria having caused serious financial loss for the country's economy. Aim: This study was undertaken to isolate and confirm the presence of M. mycoides subsp. mycoides (Mmm) in cattle, from three selected South-Eastern states of Nigeria. Method: A total of 90 bovine samples (25 pleural fluids and 65 lung tissues) suggestive of CBPP were collected from different abattoirs in the three selected South-eastern states of Nigeria (Anambra, Enugu, and Imo), for the isolation of Mmm by employing cultural method, whereas for confirmation polymerase chain reaction (PCR) approach was used. The collected samples were cultured on Pleuropneumonia like organism (PPLO) agar according to specific protocols. Results: Twenty five of the samples (lungs and pleural fluid) were positive for Mmm on PPLO agar giving an isolation rate of 27.7%. Only 21 of the isolates were further confirmed using PCR. The PCR amplification of the isolates produced a product of 1.1 kbp which is specific for Mmm. No positive isolates were recovered from Imo state. Conclusion: This study confirms the presence of Mmm as the causative organism of CBPP in Southeast Nigeria. It is recommended that active surveillance and vaccination protocol should be undertaken in the region for the control and prevention of this disease.

Salmonellosis: Serotypes, prevalence and multi-drug resistant profiles of Salmonella enterica in selected poultry farms, Kwara State, North Central Nigeria.

Salmonellosis is a major threat facing the poultry industry globally. This study was conducted to investigate the level of Salmonella contaminations and determine the resistance pattern of isolates obtained from selected poultry farms in Kwara State, a transition state between southern and northern regions of Nigeria. A total of 900 samples were collected between January and August 2017, from the poultry environment, apparently including healthy and dead birds. Salmonella was isolated and identified using standard bacteriological methods. All presumptive Salmonella isolates were serotyped and tested for antimicrobial susceptibility using 11 different antimicrobials. A total of 58 (6.4%) Salmonella isolates were obtained, and the isolation rate was only statistically significant (p < 0.05) in live birds. The isolates comprised of 13 serovars. The three predominant serovars, Salmonella enterica ser. 6.7:d:- (29.0%), Salmonella Agama (28.0%) and Salmonella Typhimurium (16.0%), were isolated from all three sample types. Rare serovars like Salmonella Albany, Salmonella Colindale, Salmonella Istanbul, Salmonella Larochelle, Salmonella Nigeria and Salmonella Orion were also isolated in this study. A high frequency of resistance was generally observed with all the isolates exhibiting a total of (100%) resistance to ampicillin, cefotaxime and ceftazidime. This study documents the first predominant isolation of S. enterica ser. 6.7:d:- and S. Agama from chickens. It also documents the high frequency of fluoroquinolone and cephalosporins resistance of the isolates indicating the presence of selective pressure in the environment. Controls and targeted interventions against Salmonella and the frequent occurrence of antimicrobial resistance in chickens should be initiated to prevent the spread of this organism.

Molecular characterization of dermatophytes isolated from cattle in Plateau State, Nigeria.

Dermatophytes from cattle were successfully characterized to species and strain levels for the first time in Nigeria. This study was undertaken to isolate and characterize dermatophytes from cattle in Plateau State, Nigeria. Two molecular techniques were utilized. The first was the use of polymerase chain reaction (PCR) to amplify the internal transcribed spacer regions of the ribosomal DNA using ITS-1 and ITS-4 as primers. This was followed by restriction fragment length polymorphism analysis of the amplified ITS regions using the enzyme MvaI to identify dermatophyte species. The second technique was a PCR using the short oligonucleotide 5'-GACAGACAGACAGACA-3' as primer for the RAPD typing of the isolates for identification of dermatophytes based on species specific profiles. Profiles of dermatophytes and their correlation with location, site of infection and severity of disease were also investigated. Both PCR-RFLP and RAPD analysis identified 26 Trichophyton verrucosum and 22 Trichophyton mentagrophytes. The PCR-RFLP analysis of the ITS regions produced two distinct profiles for both T. mentagrophytes and T. verrucosum. The first Profile for T. mentagrophytes consisted of two fragments of approximately 320 bp and 280 bp in length while the second was approximately 350 bp and 250 bp in length. The first profile for T. verrucosum consisted of two fragments having bands of approximately 380 bp and 220 bp. The second profile had a single band of undigested fragment of approximately 600 bp in length. Both T. mentagrophytes and T. verrucosum yielded identifiable fragments by RAPD analysis. Six profiles were produced for T. mentagrophytes and the PCR finger prints ranged from 1 to 9 bands with sizes ranging from approximately 350 to 5000 base pairs in size. Amplification of T. verrucosum isolates produced four Profiles. The PCR fingerprints ranged from 5 to 7 bands with sizes ranging from 500 bp-5000 bp. The results indicate that differences in location could contribute to variations in PCR amplicons of dermatophytes and strain differences in dermatophytes may be responsible for variation in clinical dermatophytosis but no significant association was observed between profiles of dermatophytes and the site of infection. The PCR-RFLP analysis of the internal transcribed spacer regions using the primer set ITS1/ITS4 and RAPD analysis using (GACA)4 as primer were successfully used to accurately identify dermatophytes from cattle to species and strain levels. Few molecular studies targeting dermatophytes of cattle are available in the literature. As far as we know, this may be the first report of molecular characterization of cattle dermatophytes from Africa.

Characterization of high level ampicillin- and aminoglycoside-resistant enterococci isolated from non-hospital sources.

PURPOSE: High level ampicillin- and aminoglycoside-resistant enterococci are being increasingly reported from non-hospital sources. This study was carried out to characterize these strains from non-hospital sources in Nigeria. METHODOLOGY: A collection of Enterococcus faecium isolated from vegetables, soil, farm animals and manure and observed to be resistant to ampicillin (n=63) and gentamicin (n=37) discs, were screened for resistance to high levels of ampicillin and aminoglycoside using E-test strips. Putative high level ampicillin- and aminoglycoside-resistant strains were screened for pbp5 and aminoglycoside modifying enzyme genes, respectively, by PCR. The C-terminal region of the amplified pbp5 gene was also sequenced. RESULTS: Five (5/63) and thirty-five (35/37) of the ampicillin- and aminoglycoside-resistant strains were identified as high level ampicillin- and aminoglycoside-resistant E. faecium strains, respectively, based on the MIC results. The amplified pbp5 gene from the high level ampicillin-resistant isolates displayed 96-99 % nucleotide sequence similarity with the reference strains and three novel insertions (500Glu→Leu, 502Asp→Arg and 614Ile→Phe) in the amino acid sequence. Aminoglycoside modifying enzyme genes aac(6')-Ie-aph(2″) (100 %), aph(2')-Ic (88.8 %), aph(3')-IIIa (90 %) and ant(4')-Ia (40 %) were detected among the high level aminoglycoside-resistant isolates. CONCLUSION: This is the first report on the characterization of high level ampicillin- and aminoglycoside-resistant Enterococcus faecium among animals and vegetables in Nigeria. The results show that non-hospital sources can constitute a reservoir for potential dissemination of these strains and genes to humans via the food chain or by direct contact.

Equine Dermatophytosis: A Survey of Its Occurrence and Species Distribution among Horses in Kaduna State, Nigeria.

This study was designed to determine the occurrence and species distribution of dermatophyte from cutaneous skin lesions of horses in Kaduna State, Nigeria. A total of 102 skin scrapings were collected from 102 horses with skin lesions. Mycological studies were carried out using conventional techniques. Dermatophytes were isolated from 18 (17.6%) of the 102 samples collected. The 18 dermatophytes were distributed into 10 different species belonging to Microsporum (n = 5) and Trichophyton (n = 5) genera. T. verrucosum (n = 4) was the most predominant species isolated followed by M. equinum (n = 3), T. vanbreuseghemii (n = 2), M. gypseum (n = 2), and M. canis (n = 2). Others include M. fulvum (n = 2), T. mentagrophytes (n = 1), T. equinum (n = 1), T. soudanense (n = 1), and M. gallinae (n = 1). The present study reveals the occurrence of dermatophytes in cutaneous skin lesions of horses in Kaduna State, Nigeria. In addition for the first time in this environment the anthropophilic dermatophyte T. soudanense was isolated from horses. These findings have great economic, veterinary, and public health significance as they relate to the cost of treatment and dissemination of zoonotic dermatophytes.

Serological prevalence of leptospirosis in cattle slaughtered in the Zango abattoir in Zaria, Kaduna State, Nigeria.

Leptospirosis is an occupational zoonosis caused by pathogenic leptospires. In this study, the presence and prevalence of antibodies specific to Leptospira spp. serovar Hardjo in 142 cattle slaughtered between June and July 2011 was investigated using the enzyme-linked immunosorbent assay (ELISA). Five (3.50%) of the 142 cattle sampled were seropositive for antibodies to Leptospira spp. serovar Hardjo. Despite the fact that there was no significant difference (p>0.05) in seropositivity between sexes and between breeds sampled, there was a significant difference (p<0.05) in sero-positivity between the different age groups examined. Leptospirosis is present in cattle slaughtered in the Zango abattoir; butchers and abattoir workers are exposed to infected animals and are at risk of being infected by Leptospira spp. serovar Hardjo.