Staphylococcus aureus in Horses in Nigeria: Occurrence, Antimicrobial, Methicillin and Heavy Metal Resistance and Virulence Potentials.

Staphylococcus aureus was isolated from a total of 360 nasal and groin skin swabs from 180 systematic randomly-selected horses slaughtered for meat at Obollo-Afor, Enugu State, Southeast Nigeria and antimicrobial, methicillin and heavy metal resistance profile and virulence potentials of the isolates established. Baird-Parker agar with egg yolk tellurite was used for S. aureus isolation. S. aureus isolates were confirmed biochemically and serologically using a specific S. aureus Staphytect Plus™ latex agglutination test kit. The antimicrobial resistance profile, methicillin, vancomycin and inducible clindamycin resistance, and ß-lactamase production of the isolates were determined with disc diffusion. Tolerance to Copper, Cadmium, Lead and Zinc was assessed using the agar dilution method and virulence potentials were determined using phenotypic methods. Forty-three (23.9%) of the 180 horses harbored S. aureus. Some 71 S. aureus were recovered from the 360 samples. Two (2.8%) of the 71 S. aureus were methicillin-resistant S. aureus (MRSA) and 69 (97.2%) were methicillin-susceptible. MRSA was recovered from 2 (1.1%) of the 180 horses. Some 9.4% of the isolates were multiple drug-resistant (MDR). The mean multiple antibiotic resistance indices (MARI) for the isolates was 0.24. Heavy metal resistance rate of the isolates ranged between 35.4-70.4%. The isolates, including the MRSA strains, displayed virulence potentials as clumping factor and catalase, gelatinase, caseinase, heamolysin, and biofilm was at the rate of 100%, 53.5%, 43.7%, 18.3% and 23.9%, respectively. This study showed that a considerable percentage of horses slaughtered in Obollo-Afor Southeastern Nigeria are potential reservoirs of virulent multiple drug- and heavy metal-resistant S. aureus, including MRSA, that could spread to humans and the environment.

Computational Evaluation of Azadirachta indica-Derived Bioactive Compounds as Potential Inhibitors of NLRP3 in the Treatment of Alzheimer's Disease.

BACKGROUND: The development of therapeutic agents against Alzheimer's disease (AD) has stalled recently. Drug candidates targeting amyloid-ß (Aß) deposition have often failed clinical trials at different stages, prompting the search for novel targets for AD therapy. The NLRP3 inflammasome is an integral part of innate immunity, contributing to neuroinflammation and AD pathophysiology. Thus, it has become a promising new target for AD therapy. OBJECTIVE: The study sought to investigate the potential of bioactive compounds derived from Azadirachta-indica to inhibit the NLRP3 protein implicated in the pathophysiology of AD. METHODS: Structural bioinformatics via molecular docking and density functional theory (DFT) analysis was utilized for the identification of novel NLRP3 inhibitors from A. indica bioactive compounds. The compounds were further subjected to pharmacokinetic and drug-likeness analysis. Results obtained from the compounds were compared against that of oridonin, a known NLRP3 inhibitor. RESULTS: The studied compounds optimally saturated the binding site of the NLRP3 NACHT domain, forming principal interactions with the different amino acids at its binding site. The studied compounds also demonstrated better bioactivity and chemical reactivity as ascertained by DFT analysis and all the compounds except 7-desacetyl-7-benzoylazadiradione, which had two violations, conformed to Lipinski's rule of five. CONCLUSION: In silico studies show that A. indica derived compounds have better inhibitory potential against NLRP3 and better pharmacokinetic profiles when compared with the reference ligand (oridonin). These compounds are thus proposed as novel NLRP3 inhibitors for the treatment of AD. Further wet-lab studies are needed to confirm the potency of the studied compounds.

Occurrence of Trypanosoma spp. and piroplasm infections of horses at Obollo-Afor southeastern Nigeria and resistance profiles of trypanosomes to isometamidium and diminazene salts.

Epidemiological data on trypanosomosis and piroplasmosis of horses are lacking in southeastern Nigeria. The prevalence of trypanosome and piroplasm infections in horses and resistance profile of isolated trypanosomes to diminazene and isometamidium salts were investigated. For the cross-sectional study of horses billed for slaughter, 304 horses were randomly sampled. Approximately 2 ml of blood was collected into anticoagulant-treated bottles for haematocrit (HCT) determination, direct microscopic examinations, and rat inoculation. Gender, body condition scores (BCS), age groups, and body weights of sampled horses were noted. Two isolates of Trypanosoma brucei recovered from the cross-sectional study were profiled for resistance to isometamidium hydrochloride and diminazene diaceturate in 36 BALB/c mice. Standardized protocols were used (Eisler et al., Veterinary Parasitology 97:171-182, 2001). 19.1% of horses (95% confidence interval 14.7-23.5%) were positive for haemoparasite infections including Theileria equi (16.1%) and Babesia caballi (3.9%). Only two (0.66%) Trypanosoma brucei infections were seen, being from active cases. Associations between age or gender, and presence of haemoparasites were only random. Haemoparasite-infected horses had significantly (p < 0.05) lower mean HCT and body weights and poorer BCS. From resistance profiling, for each isolate, all mice in control groups were parasitaemic by day 6 post-inoculation, while mice in test groups remained aparasitaemic over 60-day observation period. The study showed the endemicity and weights of Trypanososma spp. and piroplasm infections and among horses within the area. Furthermore, circulating strains of Trypanosoma brucei in the area are still susceptible to isometamidium and diminazene salts in mice. The pharmacoepidemiological significances of these findings were discussed.

Hematobiochemical and Immunological Responses of Rats Treated with Multi-strain Probiotics and Infected with Trypanosoma brucei.

The effects of treatment with probiotics on the immunological and hematobiochemical changes in Trypanosoma brucei infection were investigated. Probiotic strains used are Bifidobacterium BB-12, Lactobacillus acidophilus LA-5, Lactobacillus delbrueckii LBY-27, Lactobacillus paracasei LC-01, and Streptococcus thermophilus STY-31. Thirty rats randomly assigned to five groups were used in the experiment. Groups A to C received 1 × 109 CFU, 5 × 109 CFU, and 10 × 109 CFU of the multi-strain probiotics daily and respectively from day 0 post-supplementation (PS) to termination. Group D and E were the infected and uninfected controls respectively. On day seven PS, groups A to D were challenged intraperitoneally with approximately 1 × 106 trypanosomes. Parasitemia, nitric oxide level, hematobiochemical parameters, and antibody titer to heterologous antigen stimulation were monitored post-infection. By days 7 and 16 PS, probiotics-treated groups had significantly lower (p < 0.05) mean creatinine concentration than the controls; however, on day 7 PS, there were no significant variations in the leukocyte counts (LC), total erythrocyte counts (TEC), and the packed cell volume (PCV) in all experimental groups. Following infection, by day 16 PS, the pre-patent period, parasitemia levels, and antibody titer were similar in all infected groups. Furthermore, the probiotics-treated groups and the infected control had significantly lower PCV, TEC, and LC values when compared to the uninfected control, and probiotics treated groups (A and C) had only marginally lower nitric oxide levels than the infected control. Treatment with the probiotic strains gave a creatinine-lowering effect, was innocuous to the hematopoietic system, but was not sufficiently immunostimulatory in trypanosomosis.