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This study was conducted to determine whether there exists the presence of plasmid from Salmonella isolated from poultry farms in Abia and Imo states southeast Nigeria. Forty Salmonella isolates were used for the study. The alkaline phosphate method of Birnboin and Doly was employed. Three buffers A, B and C made up of different concentrations and volumes with adjusted pH were used. A (400 mM Tris, 200 mM Na Ethylenediaminetetraacetic acid), B (3 M Na, acatic acid) and C (10 mM Tris, 2 M Na Ethylenediaminetetraacetic acid). The test organisms sub-cultured on MacConkey agar (MCA) were processed at various centrifugation rates and time. The suspected pure plasmid deoxyribonucleic acid was mixed with ethidium bromide and loading dye using agarose gel. The set up was viewed under ultraviolet transillumination with gel documentation apparatus. Plasmid cure was conducted with ten multidrug resistant Salmonella isolates. Danifor Biotechnology method was used for the plasmid curing experiment. Three curing agents, sodium deodecyl sulphate curing agents (100), sodium deodecyl sulphate (10 g or 10%) and nutrient broth (100 ml) were used. Plasmid DNA was detected from the Salmonella isolates evidenced with bands at 100 bp. There was plasmid cure as supported by the zones of inhibition by some of the antibiotics when compared with the original isolates.Keywords: Cure, DNA, multidrug resistant isolates, plasmid, Salmonella

Objectives Resistance to colistin (CST) mediated by mobile genetic elements has had a broad impact worldwide. There is an intensified call for epidemiological surveillance of mcr in different reservoirs to preserve CST for future generations. In Nigeria, the poultry industry is a key livestock sector. This study was undertaken to screen putative colistin-resistant Enterobacterales (CST-r-E) from poultry birds in Southeast Nigeria and to determine the genetic relatedness of mcr-harbouring isolates. Methods Faecal and cloacal swab samples (n = 785) were collected from chickens in 17 farms located in three contiguous states in Southeast Nigeria between March–November 2018. Following selective culture, CST-r-E were isolated. Confirmation of CST resistance, antimicrobial susceptibility testing, molecular detection of genes mcr-1 to mcr-10, multilocus sequence typing (MLST) and randomly amplified polymorphic DNA (RAPD) analysis were performed on the isolates. A questionnaire was distributed to investigate the knowledge about CST and its use of chicken farm caretakers. Results Of the 785 samples evaluated, 45 (5.7%) were positive for 48 CST-r-E, among which 23 harboured the mcr-1 gene (22 Escherichia coli and 1 Klebsiella pneumoniae). In two E.coli isolates, a new allelic variant (mcr-1.22) was detected. RAPD analysis allowed the identification of 11 different fingerprints. MLST also revealed 11 STs, with 3 of them being novel. Conclusion mcr has significantly spread in poultry birds of Southeast Nigeria, which poses a worrisome risk to veterinary and human health. Strategies to prevent indiscriminate use of CST in farms should be quickly adopted before CST resistance becomes a huge global health issue.

Synergistic interaction of adsorbents in reducing the adverse impacts of mycotoxin on performance and proximate composition of broiler feeds was investigated. Fungal growth was induced by sprinkling water on the feed. S. cerevisiae + bentonite, kaolin + bentonite or S. cerevisiea + kaolin adsorbent combinations (1.5 g/kg feed) were added and the feeds were stored in black polythene bags. An untreated group was kept as a positive control while fresh uncontaminated feed was used as a negative control. Mycotoxins were extracted from the feeds and quantified using reverse phase HPLC. Proximate composition, nutrient digestibility of the feeds, feed intake and weight gain of the broilers were measured. Deoxynivalenol (DON) concentration in the contaminated/untreated feed was 347 µg/kg while aflatoxin B1 (AFB1) was 34 µg/kg. Addition of bentonite and kaolin in the contaminated feed reduced AFB1 and DON to significantly lower levels. Feed intake and weight gain were low in the broilers fed the contaminated feed. The carbohydrate level was significantly (p &lt, 0.05) reduced from 62.31 to 40.10%, crude protein digestibility dropped from 80.67 to 49.03% in the fresh feed and contaminated feed respectively. Addition of the adsorbents (S. cerevisiae and bentonite) significantly (p &lt, 0.05) improved these parameters.

Antimicrobial resistance is a growing public health problem and a threat to effective treatment and prevention of an array of infections caused by bacteria. Africa is already faced with many socio-economic and health crises. Many countries in Africa can seldom boast of a standardized health care facility comparable to those in developed countries. Yet, the non-therapeutic use of COL has been banned in developed countries. However, in Africa, except for South Africa, COL is an over-the-counter (OTC) medication sold and dispensed by non-professionals/without a veterinarian’s supervision. The ban of non-therapeutic COL in developed countries has proven to reduce the development of mobile colistin resistance (MCR) in humans and animals. The unregulated use of COL has been proven to select pathogenic and commensal bacteria resistance. A transmissible plasmid-mediated colistin determinant, mobile COL resistance (mcr) gene, which is rapidly transferred/acquired horizontally or laterally intra/inter-species/genera, has been reported. A highly promiscuous mobile genetic element like plasmids containing transposons, insertion sequences, and integrons aid the carriage/rapid transfer and acquisition of these mcr genes. Hence, we highlight the danger posed by escalating colistin (COL) resistance in the continent and the impetus to halt the indiscriminate and non-therapeutic use of COL to protect public health.

Idiopathic pulmonary arterial hypertension (IPAH) is a rare but fatal disease diagnosed by right heart catheterisation and the exclusion of other forms of pulmonary arterial hypertension, producing a heterogeneous population with varied treatment response. Here we show unsupervised machine learning identification of three major patient subgroups that account for 92% of the cohort, each with unique whole blood transcriptomic and clinical feature signatures. These subgroups are associated with poor, moderate, and good prognosis. The poor prognosis subgroup is associated with upregulation of the ALAS2 and downregulation of several immunoglobulin genes, while the good prognosis subgroup is defined by upregulation of the bone morphogenetic protein signalling regulator NOG, and the C/C variant of HLA-DPA1/DPB1 (independently associated with survival). These findings independently validated provide evidence for the existence of 3 major subgroups (endophenotypes) within the IPAH classification, could improve risk stratification and provide molecular insights into the pathogenesis of IPAH., Idiopathic pulmonary arterial hypertension is a rare and fatal disease with a heterogeneous treatment response. Here the authors show that unsupervised machine learning of whole blood transcriptomes from 359 patients with idiopathic pulmonary arterial hypertension identifies 3 subgroups (endophenotypes) that improve risk stratification and provide new molecular insights.

Plant phenolic compounds (phenylpropanoids) act as defense chemicals against herbivores and can mediate ecosystem processes. Tropospheric ozone (O

Climate warming can influence interactions between plants and associated organisms by altering levels of plant secondary metabolites. In contrast to studies of elevated temperature on aboveground phytochemistry, the consequences of warming on root chemistry have received little attention. Herein, we investigated the effects of elevated temperature, defoliation, and genotype on root biomass and phenolic compounds in trembling aspen (Populus tremuloides). We grew saplings of three aspen genotypes under ambient or elevated temperatures (+4-6 °C), and defoliated (by 75%) half of the trees in each treatment. After 4 months, we harvested roots and determined their condensed tannin and salicinoid (phenolic glycoside) concentrations. Defoliation reduced root biomass, with a slightly larger impact under elevated, relative to ambient, temperature. Elevated temperature decreased condensed tannin concentrations by 21-43% across the various treatment combinations. Warming alone did not alter salicinoid concentrations but eliminated a small negative impact of defoliation on those compounds. Graphical vector analysis suggests that effects of warming and defoliation on condensed tannins and salicinoids were predominantly due to reduced biosynthesis of these metabolites in roots, rather than to changes in root biomass. In general, genotypes did not differ in their responses to temperature or temperature by defoliation interactions. Collectively, our results suggest that future climate warming will alter root phytochemistry, and that effects will vary among different classes of secondary metabolites and be influenced by concurrent ecological interactions such as herbivory. Temperature- and herbivory-mediated changes in root chemistry have the potential to influence belowground trophic interactions and soil nutrient dynamics.

The epidemiology of Staphylococcus aureus in food animals, associated products, and their zoonotic potential in Nigeria are poorly understood. This study aimed to provide data on the prevalence, genetic characteristics and antimicrobial resistance of S. aureus isolated from chicken and pig carcasses, and persons in contact with the carcasses at slaughterhouses in Nigeria. Surface swabs were collected randomly from 600 chicken and 600 pig carcasses. Nasal swabs were collected from 45 workers in chicken slaughterhouses and 45 pig slaughterhouse workers. S. aureus isolates were analyzed by spa typing. They were also examined for presence of the Panton-Valentine Leucocidin (PVL) and mecA genes, as well as for antimicrobial resistance phenotype. Overall, 53 S. aureus isolates were recovered (28 from chicken carcasses, 17 from pig carcasses, 5 from chicken carcass handlers and 3 from pig carcass handlers). Among the isolates, 19 (35.8%) were PVL-positive and 12 (22.6%) carried the mecA gene. The 53 isolates belonged to 19 spa types. The Based Upon Repeat Pattern (BURP) algorithm separated the isolates into 2 spa-clonal complexes (spa-CC) and 9 singletons including 2 novel spa types (t18345 and t18346). The clonal complexes (CC) detected were CC1, CC5, CC8, CC15, CC88 and CC152. CC15-related isolates represented by spa type t084 (32.1%) and CC5 represented by spa type t311 (35.3%) predominated among isolates from chicken carcasses/ handlers, and pig carcasses/ handlers, respectively. Multidrug resistance exhibited by all the CC except CC8, was observed among isolates from chicken carcasses (64.3%), pig carcasses (41.2%), handlers of chicken meat (40.0%) and handlers of pork (33.3%). All the CC showed varying degrees of resistance to tetracycline while CC15 and CC5 exhibited the highest resistance to sulphamethoxazole/trimethoprim and erythromycin, respectively. The predominant antimicrobial resistance pattern observed was penicillin-tetracycline-sulphamethoxazole/trimethoprim (PEN-TET-SXT). In conclusion, food animals processed in Enugu State in Southeast Nigeria are potential vehicles for transmission of PVL-positive multiple-drug resistant S. aureus and methicillin-resistant S. aureus from farm to slaughterhouse and potentially to the human population. Public health intervention programs at pre- and post-slaughter stages should be considered in Nigerian slaughterhouses.

Peste des petits ruminants (PPR) is a highly contagious, trans-boundary viral disease of sheep and goats that have hindered successful small ruminant farming. Its current status in South East Nigeria with respect to its prevalence and farmers’ awareness was studied. Three states, Anambra, Ebonyi, and Enugu, were randomly selected for the study. Sera samples from 113 goats and 172 sheep (collected from December 2017 to June 2018) were randomly collected and analysed for the presence of PPRV antibodies, while structured interview schedules were conducted to elicit information on farmers’ awareness of the disease and PPR vaccination and use of veterinary services. An overall seroprevalence of 42.5% (121/285) was recorded. The seroprevalence in decreasing order was 62.2% (Enugu), 34.8% (Anambra) and 20.3% (Ebonyi). There was a significant association (X2 = 36.08, df = 2, p = 0.0001) between seroprevalence and the state sampled. Lack of awareness of PPR vaccination among small ruminant farmers, their limited use of veterinary services (38% consult veterinarians) and non-availability of the vaccine at veterinary establishments in the sampled states are potential risk factors of PPR prevalence in South East Nigeria. Consequently, an effective control measure like mass vaccination is recommended for the study area. Also, there is a need for an extension program for stakeholders and farmers in the study area and country on the grave importance and economic benefits of PPR vaccination and the use of veterinary services.

Aim An epidemiological surveillance for Staphylococci contamination of ready-to-eat (RTE) meats from Enugu State, Nigeria, was carried out to determine the prevalence, species distribution, toxigenic potential and antimicrobial susceptibility profile of the organisms and hence the microbiological and toxicological safety of the meats. Materials and methods Isolation and phenotypic Staphylococcus detection were done according to standard microbiological methods. Phenotypic resistance to 17 commonly used antimicrobial agents was determined by disc diffusion method. Molecular characterization of the isolates to species level and detection of selected toxigenic and antimicrobial-resistance genes were done by PCR methods. Results Twenty-four (9.4%) of the 255 meat samples investigated were contaminated with Staphylococcus species. Twenty-four Staphylococcus isolates belonging to six species of coagulase-negative Staphylococcus (CoNS) were identified. Four (16.7%) isolates harbored genes coding for exfoliative toxin-A. Ten (41.7%) isolates were multidrug resistant, while mecA, tetK, mphC, ermT and ermC were the antimicrobial-resistance genes detected in the isolates. Meat samples sourced from motor parks (16.7%) and open markets (8.5%) were the most contaminated. Conclusion 9.4% of RTE meats sampled were contaminated with toxigenic and multidrug resistance CoNS. Beef was the most contaminated RTE meat type and harbored all the toxigenic and most of the antibiotic-resistant genes detected. Meat samples from motor parks had the highest staphylococcal contamination (16.7%), while those from mechanic village had the least (2.4%). Majority (79.2%) of the isolates were not susceptible to fusidic acid but none exhibited antimicrobial-resistance to chloramphenicol, ciprofloxacin, linezolid or teicoplanin. Food safety authorities in the study area should work proactively to massively improve the hygienic practices of meat vendors; in order to limit staphylococcal contamination of RTE meats and the associated public health problems.

Oral fowl typhoid (FT) vaccine is necessary for improved flock vaccinations and economic growth. This study was undertaken to evaluate the immune responses of birds given oral fowl typhoid vaccine coated with chitosan/alginate microparticles and comparing it with the conventional subcutaneous route of administration. Preliminary studies were done to evaluate the particle size, encapsulation efficiency and agglutination. Sixty day-old chicks were divided into three groups of twenty birds each. This comprised a negative control group NEG 451 (non-vaccinated and non-challenged used as control for cytokine quantification), SC 634 (live 9R vaccine by the injection route) and OCV 567 (live 9R vaccine coated with chitosan/alginate microparticles). Vaccination was done at 10 weeks and 14 weeks of age followed by challenge at 16 weeks of age. IgG was measured using ELISA. mRNA fold expression of IFN-γ in spleen was calculated using qRT-PCR. Particle sizes ranged between 0.55 µm and 10 µm. Encapsulation efficiency was above 60%. ELISA showed E-values of 0.10 ± 0.14, 0.07 ± 0.01 and 0.02 ± 0.01 for OCV 567, SC 634 and NEG 451 respectively after primary vaccination. Also E-values were 0.25 ± 0.16, 0.19 ± 0.04 and 0.0008 ± 0.005 for SC 634, OCV 567 and NEG451 respectively after boost vaccination. The expression of IFN-γin spleen using 2-ΔΔ CT calculation was upregulated with values of 1.97 and 0.75 for OCV 567 and SC 634 respectively. After challenge with the 85-kb virulence plasmid SG9, there was 100% protection of the birds in both OCV 567 and SC 634 groups with no mortality. In conclusion, there was no significant difference at p < 0.05 of the means ± SD in immune responses between the oral fowl typhoid vaccine coated with chitosan/alginate microparticles and the subcutaneous route of administration. However, it is noteworthy to mention that the protective efficacy of the oral route is due to the chitosan/alginate biopolymers which coated the vaccine preventing destruction in the gastrointestinal tract.

Synthesis, electronic, infrared, elemental micro analytical studies were carried on N-(benzothiazol-2-yl)trichloroethanamide [4] and N-(benzothiazol-2-yl)chloroethanamide [5]. They were also screened in vitro and in vivo for antibacterial activity. The results indicate that the compounds are very stable and that they show high antibacterial activities against both gram-positive and gram-negative bacteria tested. Both derivatives of 2-aminobenzothiazole were active against the multiresistant bacteria with IZD ranging from 9 -18 mm [5] and 9 - 20mm [4]. From the MIC results it is observed that the [5] derivative produced a better antibacterial activity than the [4] derivative. The lethal concentrations (LC50) of the compounds were also determined. Their solubilities and melting points were also determined.

The study was conducted at Obollo-Afor horse market to determine the occurrence and the antifungal susceptibility profile of mycotic agents in skin lesions of trade horses. The samples were collected by plucking the hair and scraping the epidermal scales using sterile scapel blades. Each sample was planted onto the surface of Sabouraud dextrose agar (SDA) media plate. Inoculated plates was incubated at 25–30 °C and examined daily for fungal growth for 4 weeks. After obtaining pure cultures, they were subcultured on the Potato dextrose agar (PDA) for proper sporulation. Identification of each of the fungal isolates were based on cultural characteristics, microscopic examinations, slide culture technique and biochemical tests. 150 (30.18%) of the examined horses had skin lesions, 97 (64.7%) of which were males while 53 (35.3%) were females. In the wet season, 140 (93.3%) of the horses with skin lesions were observed and 10 (6.6%) were observed to have lesions during the dry season. The overall isolation rates were as follows the genera Aspergillus (24.5%), Trichophyton (14.3%), Microsporum (6.1%), Penicillium (8.2%), Curvularia (6.1%) and Fusarium (4.1%) from skin lesions of horses. All the isolates apart of those of Penicillium spp. were resistant to fluconazole while none of the respective isolates were resistant to voriconazole and nystatin. This study has provided baseline information which includes sex and seasonal distributions of the fungal agents associated with skin lesions in trade horses in Obollo-Afor area of Enugu State. Antifungal susceptibility results shows that voriconazole and nystatin are the drugs of choice in the treatment of dermatomycoses of horses in Nigeria.

Survival of Ammopiptanthus mongolicus in a cold desert environment is facilitated by high photosynthesis rates in spring and summer, and efficient photoprotective strategies in winter cold. Woody evergreen plants inhabiting cold desert ecosystems must retain their foliage amidst chronically dry conditions and large seasonal temperature variations. To understand the strategies enabling survival of evergreens in these environments, we monitored seasonal changes in foliar gas exchange and photosynthetic traits of Ammopiptanthus mongolicus, an evergreen broadleaved shrub native to the cold desert of northwestern China. We found that photosynthesis was relatively higher in spring and summer and lower in fall and winter. Transitioning from spring to summer, A. mongolicus maintained high photosynthetic capacity (Amax). Transitioning into fall, the Amax and maximum stomatal conductance (gsmax) decreased, while the relative stomatal limitation to photosynthesis (Ls) increased. In winter, A. mongolicus decreased Amax, maximum quantum efficiency of photosystem II (Fv/Fm), maximum RuBisCo carboxylation rates (Vcmax), maximum RuBP regeneration rates (Jmax), and photosynthetic nitrogen-use efficiency (PNUEmax) relative to other seasons. Collectively, these results suggest that A. mongolicus adapts physiologically to maximize carbon assimilation during spring and summer, and to maximize foliar resistance to cold stress at the expense of photosynthesis in winter. Foliage was protected against photo-oxidative damage during temperature extremes in winter by dark-sustained thermal energy dissipation. Overall, our study reveals that multiple photosynthetic adjustments, varying among the seasons, enable the survival of cold desert evergreens.

Understanding the sources, prevalence, phenotypic and genotypic characteristics of mcr gene-harbouring bacteria (MGHB) in the poultry sector is crucial to supplement existing information. Through this, the plasmid-mediated colistin resistance (PMCR) could be tackled to improve food safety and reduce public health risks. Therefore, we conducted a literature synthesis of potential sources and characteristic occurrence of MGHB recovered from the poultry sector specific to the high-income countries (HICs). Colistin (COL) is a last-resort antibiotic used for treating deadly infections. For more than 60 years, COL has been used in the poultry sector globally, including the HICs. The emergence and rapid spread of mobile COL resistance (mcr) genes threaten the clinical use of COL. Currently, ten mcr genes (mcr-1 to mcr-10) have been described. By horizontal and vertical transfer, the mcr-1, mcr-2, mcr-3, mcr-4, mcr-5, and mcr-9 genes have disseminated in the poultry sector in HICs, thus posing a grave danger to animal and human health, as harboured by Escherichia coli, Klebsiella pneumoniae, Salmonella species, and Aeromonas isolates. Conjugative and non-conjugative plasmids are the major backbones for mcr in poultry isolates from HICs. The mcr-1, mcr-3 and mcr-9 have been integrated into the chromosome, making them persist among the clones. Transposons, insertion sequences (IS), especially ISApl1 located downstream and upstream of mcr, and integrons also drive the COL resistance in isolates recovered from the poultry sector in HICs. Genes coding multi-and extensive-drug resistance and virulence factors are often co-carried with mcr on chromosome and plasmids in poultry isolates. Transmission of mcr to/among poultry strains in HICs is clonally unrestricted. Additionally, the contact with poultry birds, manure, meat/egg, farmer’s wears/farm equipment, consumption of contaminated poultry meat/egg and associated products, and trade of poultry-related products continue to serve as transmission routes of MGHB in HICs. Indeed, the policymakers, especially those involved in antimicrobial resistance and agricultural and poultry sector stakeholders-clinical microbiologists, farmers, veterinarians, occupational health clinicians and related specialists, consumers, and the general public will find this current literature synthesis very useful.

Variety of biological products has been applied to stimulate biologic factors and promote osteogenesis. In spite of this, surgeons are still facing challenges relating to fracture healing. The methods of preparation of these biologic factors influence their efficacy and should be evaluated and understood prior to their use in clinical trials. The use of batroxobin in platelet-rich plasma activation has been documented. This study was designed to evaluate the osteogenic effects of two platelet-rich plasmas prepared using different activating agents (batroxobin/calcium gluconate and calcium chloride alone) on tibia fractures of Nigerian local dogs. Nine healthy Nigerian local dogs (males) were used for the study. They were randomly assigned into three groups, viz, Plateltex-prepared platelet-rich plasma-treated (PPRP) group (n = 3); calcium chloride-activated platelet-rich plasma-treated (CPRP) group (n = 3); and normal saline-treated control group (CG), (n = 3). Transverse fractures were created aseptically on the left tibia of each animal under general anesthesia. The fractures were reduced and immobilized in a retrograde fashion using Steinmann intramedullary pins. Before closure, the PPRP fractures were treated with batroxobin/calcium gluconate-activated autologous PRP; CPRP fractures with calcium chloride-activated autologous PRP while the normal saline-treated control fractures were treated with normal saline. Radiographs of the fractures were taken at weeks 1, 2, 4, and 8 post treatments and the rate of healing evaluated subjectively and semi-quantitatively without bias. The PPRP dogs had better functional limb usage in terms of early weight bearing. Radiologically, there was however no statistical significant difference (p