Co-infection of Helicobacter pylori and intestinal parasites in children of selected low-income communities in Lagos State, Nigeria.

Helicobacter pylori and intestinal parasites cause gastrointestinal diseases with a high prevalence in children in resource limited developing countries. There is paucity of information in Nigeria on co-infection of H. pylori and intestinal parasites. The study was conducted to determine the prevalence of H. pylori and parasite co-infection in children from selected low-income communities in Lagos, Nigeria. Fecal samples were collected from 151 healthy children aged ≤11 years across six low-income communities in Lagos. H. pylori was detected using stool antigen test and conventional PCR assay, intestinal parasites were detected using formol-ether concentration and nested PCR assay. Structured questionnaires were administered to parents and legal guardians of the children by an interviewer to collect relevant data on demographic and lifestyle factors. The prevalence of H. pylori was 31.79% (48), with a higher prevalence in children aged 2-3 years. The prevalence of intestinal parasites was 21.19% (32) with the lowest frequency found in children aged 8-9 years. The parasites detected include: A. lumbricoides (10.6%), G. intestinalis (7.3%), hookworm (1.99%), E. histolytica (0.66%), S. mansoni (0.66%). There was co-infection prevalence of 10.6% (16) which was associated with the parasites: G. intestinalis (7.3%) and A. lumbricoides (3.97%). Polyparasitism with G. intestinalis and A. lumbricoides was reported in 2 children infected with H. pylori. This study which is the first reported in Lagos established a low prevalence of H. pylori and intestinal parasite co-infection in children and provides better understanding of the epidemiology of H. pylori infection associated with intestinal parasites in Nigeria.

Direct detection of Helicobacter pylori from biopsies of patients in Lagos, Nigeria using real-time PCR-a pilot study.

OBJECTIVE: Prompt diagnosis of Helicobacter pylori infection is essential for proper treatment and eradication of the pathogen because prolonged infection could lead to gastric cancer. Sensitive and cost effective diagnostic methods are key to guiding treatment options that will reduce mortality. This study was aimed at detecting H. pylori from biopsies of peptic ulcer patients. Real-time PCR using TaqMan and EvaGreen assays targeting 16S rRNA and ureA genes were used to detect H. pylori DNA extracted from 40 biopsy samples comprising 20 biopsies obtained from the antrum and 20 from the corpus of 20 patients undergoing endoscopy for duodenal ulcer investigation in Lagos, Nigeria. RESULTS: H. pylori was detected in 80% of the biopsy samples by combined cycle threshold (Ct) and melting temperature (Tm) values. Mean Ct value for ureA gene ranged from 21.40 to 37.53 and 22.71 to 35.44 for 16SrRNA gene. Average melting temperatures (Tm) of 81.57 and 82.90 °C among amplicons of ureA and 16S rRNA were observed respectively. H. pylori DNA was generally detected in biopsies collected from antrum and corpus. Real-time PCR in the diagnosis of H. pylori can be considered a simple, low cost and efficient alternative or addition to the gold standard.

Invited review: Sustainable forage and grain crop production for the US dairy industry.

A resilient US dairy industry will be underpinned by forage and crop production systems that are economically, environmentally, and socially sustainable. Land use for production of perennial and annual forages and grains for dairy cattle must evolve in response to multiple food security and environmental sustainability issues. These include increasing global populations; higher incomes and demand for dairy and other animal products; climate change with associated temperature and moisture changes; necessary reductions in carbon and water footprints; maintenance of soil quality and soil nutrient concerns; and competition for land. Likewise, maintaining producer profitability and utilizing practices accepted by consumers and society generally must also be considered. Predicted changes in climate and water availability will likely challenge current feed and dairy production systems and their national spatial distribution, particularly the western migration of dairy production in the late 20th century. To maintain and stabilize profitability while reducing carbon footprint, particularly reductions in methane emission and enhancements in soil carbon sequestration, dairy production will need to capitalize on genetic and management innovations that enhance forage and grain production and nutritive value. Improved regional and on-farm integration of feed production and manure utilization is needed to reduce environmental nitrogen and phosphorus losses and mitigate greenhouse gas emissions. Resilient and flexible feed production strategies are needed to address each of these challenges and opportunities to ensure profitable feeding of dairy cattle and a sustainable dairy industry.

Assessment of knowledge of participants on basic molecular biology techniques after 5-day intensive molecular biology training workshops in Nigeria.

The deployment of molecular biology techniques for diagnosis and research in Nigeria is faced with a number of challenges, including the cost of equipment and reagents coupled with the dearth of personnel skilled in the procedures and handling of equipment. Short molecular biology training workshops were conducted at the Nigerian Institute of Medical Research (NIMR), to improve the knowledge and skills of laboratory personnel and academics in health, research, and educational facilities. Five-day molecular biology workshops were conducted annually between 2011 and 2014, with participants drawn from health, research facilities, and the academia. The courses consisted of theoretical and practical sessions. The impact of the workshops on knowledge and skill acquisition was evaluated by pre- and post-tests which consisted of 25 multiple choice and other questions. Sixty-five participants took part in the workshops. The mean knowledge of molecular biology as evaluated by the pre- and post-test assessments were 8.4 (95% CI 7.6-9.1) and 13.0 (95 CI 11.9-14.1), respectively. The mean post-test score was significantly greater than the mean pre-test score (p < 0.0001). The five-day molecular biology workshop significantly increased the knowledge and skills of participants in molecular biology techniques. © 2017 by The International Union of Biochemistry and Molecular Biology, 45(4):313-317, 2017.

Typhoidal and non-typhoidal Salmonella infections in Africa.

Salmonella infections in humans can range from self-limiting gastroenteritis typically associated with non-typhoidal Salmonella (NTS) to typhoidal fever, which can be life-threatening. Salmonellosis causes considerable morbidity and mortality in both humans and animals, and has a significant socioeconomic impact worldwide. In Africa, it is difficult to evaluate the situation of salmonellosis due to the non-availability of facilities capable of performing the tests essential for the diagnosis of typhoidal and non-typhoidal Salmonella infections. This article reviews important work in the literature, including the epidemiology, disease burden, pathogenesis, genomics, diagnosis, treatment, emergence and tracking of multidrug-resistant (MDR) Salmonella infections and intercontinental transmission of Salmonella to Africa. Searches of PubMed and Google Scholar were completed and the retrieved list of relevant publications were further screened. The literature revealed that the most common form of the disease in Africa is gastroenteritis, with bacterial multiplication in intestinal submucosa and diarrhoea caused by the inflammatory response and, perhaps, also by toxins. In addition to the high burden of Salmonella infection in Africa, MDR Salmonella species is on the rise in the continent, which might pose difficulties in the treatment of the disease.

Invited review: Sustainability of the US dairy industry.

The US dairy industry has realized tremendous improvements in efficiencies and milk production since the 1940s. During this time, farm and total cow numbers have decreased and average herd size has increased. This intensification, combined with the shift to a largely urban public, has resulted in increased scrutiny of the dairy industry by social and environmental movements and increased concern regarding the dairy industry's sustainability. In response to these concerns, a group of scientists specializing in animal welfare, nutrient management, greenhouse gas emissions, animal science, agronomy, agricultural engineering, microbiology, and economics undertook a critical review of the US dairy industry. Although the US dairy system was identified as having significant strengths, the consensus was that the current structure of the industry lacks the resilience to adapt to changing social and environmental landscapes. We identified several factors affecting the sustainability of the US dairy industry, including climate change, rapid scientific and technological innovation, globalization, integration of societal values, and multidisciplinary research initiatives. Specific challenges include the westward migration of milk production in the United States (which is at odds with projected reductions in precipitation and associated limitations in water availability for cattle and crops), and the growing divide between industry practices and public perceptions, resulting in less public trust. Addressing these issues will require improved alignment between industry practices and societal values, based upon leadership from within the industry and sustained engagement with other interested participants, including researchers, consumers, and the general public.

The use of Helicobacter pylori stool antigen test for the diagnosis of Helicobacter pylori in Lagos, Nigeria.

OBJECTIVES: This study was carried out to screen the use of Helicobacter pylori stool antigen (HpSA) tests for diagnosis and monitoring of H pylori in Nigeria. METHODS: Seven hundred and forty participants were enrolled after informed consent was obtained, while 83 came back for a post-eradication test. The stool samples were taken from the patients at endoscopy and tested for HpSA. RESULTS: The proportion of patients that were positive at the pretest, 520 (70.3%) was significantly higher (Fisher's exact p = 0.001) than those positive at the post-test, 44 (53%). There was a significant difference (F = 4.106, p = 0.043) between the mean age of those that came for the pretest (40.0 +/- 14.5 years) and those that came for the post-test, 43.6 +/- 11.6 years. More males than females had the tendency to come back for a post-eradication test. CONCLUSION: Although potential bias was introduced during this study, HpSA using monoclonal antibody could still be used for diagnosis and monitoring of H pylori in Nigeria.

Detection of Mycobacterium avium subsp. paratuberculosis in skeletal muscle and blood of ewes from a sheep farm in New Zealand.

AIM: To determine whether viable Mycobacterium avium subsp. paratuberculosis (Map) is present in skeletal muscle and blood in ewes with and without Johne's disease confirmed histologically. METHODS: A total of 51 mixed-aged ewes in poor body condition from a farm with a history of clinical Johne's disease were culled and examined at necropsy. BACTEC radiometric culture was performed on samples of skeletal muscle from the biceps femoris, mononuclear cells in peripheral blood (hereafter referred to as blood), and ileum. Histological sections and Ziehl-Neelsen (ZN)-stained impression smears of terminal ileum and mesenteric lymph nodes were examined. Ewes were defined as having confirmed Johne's disease if there was histopathological evidence typical of the disease within the ileum and adjacent lymph nodes. RESULTS: Eighteen of 21 (86%) ewes with confirmed clinical Johne's disease were culture-positive for Map from sites peripheral to the alimentary tract, comprising 15 from skeletal muscle and 13 from blood. Five of 30 (17%) ewes that did not have Johne's disease were culture-positive, with four from skeletal muscle and one from blood. The likelihood that ewes with confirmed Johne's disease had systemic Map infection compared with ewes without was determined as OR=30 (95% CI=6.3-142.0; p<0.001). CONCLUSION: The prevalence of Map infection of skeletal muscle and blood in ewes with confirmed Johne's disease was 71% and 62% respectively, and in unaffected ewes was 13% for muscle and 3% for blood. CLINICAL RELEVANCE: Skeletal muscle and blood are potential sources of exposure of humans to Map, and the risk appears higher from sheep with Johne's disease.

Antibacterial activities of lactic acid bacteria isolated from selected vegetables grown in nigeria: a preliminary report

Members of lactic acid bacteria (LAB) are known probiotics and have been reported to have antimicrobial properties. Although various researchers have documented the isolation of these bacteria from fruits and vegetables; studies on LAB associated with lettuce; cucumber and cabbage are limited and non-existing in Nigeria. This study was designed to assess lettuce; cucumber and cabbage as potential sources of LAB and investigate the actions of their bacterial cell supernatants (BCS) on some pathogenic bacteria. Using standard microbiological methods; isolated LAB were identified to species level with API 50 CH kits (Biomerieux; France). Cell free supernatants (CFS) from de Man Rogosa Sharpe (MRS) broth cultures of the LAB strains were used to challenge Pseudomonas aeruginosa ATCC 27853; Enterococcus faecalis ATCC 29212; Escherichia coli ATCC 12900 and Proteus penneri ATCC 13315 by agar well diffusion method. The control consisted of the sterile MRS broth subjected to the same growth conditions as LAB broth cultures. A total of four lactic acid bacteria were isolated as follows: Pediococcus pentosaceus 2 from cucumber; Lactobacillus cellobiosus from cabbage; Lactobacillus salivarius and Lactobacillus plantarum 1 from lettuce. Pediococcus pentosaceus 2 and L. salivarius showed inhibitory effects on all the standard strains tested while L. plantarum 1 showed no inhibitory activity against E. faecalis and E. coli. Lactobacillus cellobiosus showed inhibition against all except P. penneri. Although; the molecular characterisation and probiotic potentials of these LAB strains are being investigated in an on-going study; we presumed these vegetables are prospective sources of the bacteria in Nigeria and therefore the need to extensively investigate the vegetables and other related vegetables becomes imperative

Antibiotic susceptibility patterns in Helicobacter pylori strains from patients with upper gastrointestinal pathology in western Nigeria.

A total of 186 Helicobacter pylori isolates and 532 gastric biopsies recovered from 532 patients with varying degrees of gastroduodenal pathology are subjected to in vitro antibiotic susceptibility testing using the disc-diffusion method, Etest (MIC breakpoints) and molecular testing using the polymerase chain reaction (PCR). In the isolates studied, antibiotic resistance was as follows: piperacillin (72%), amoxicillin (66%), erythromycin (78%), tetracycline (100%) and metronidazole (95%). All isolates were sensitive to ofloxacin, ciprofloxacin and norfloxacin. None of the 245 amplicons (positive for H. pylori) from the biopsies were digested with the Bbs1 and Bsal restriction enzyme used in the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, showing sensitivity to clarithromycin. However, a 238 bp fragment from H. pylori chromosomal DNA (corresponding to the quinolone resistance determining region [QRDR]) of the gyrA gene was amplified successfully. Twelve (4.9%) of the 245 strains studied had the described mutation at position 91, from asparagine (Asn) to glycine (Gly). The study showed that all the H. pylori strains were sensitive to clarithromycin and ciprofloxacin. It also highlighted PCR as a potential tool for faster diagnosis and determination of antibiotic susceptibility (within 24 h) of H. pylori from biopsies and/or isolates recovered from peptic ulcer and gastritis patients.

Antibiotic susceptibility pattern of Staphylococcus species isolated from telephone receivers.

INTRODUCTION: Microorganisms are transferred to everyday objects from the environment and infected individuals. Pathogenic microbes are transmissible from the air, skin, hands and other interpersonal contacts and cause diseases most of the time. This study centres on the microbial assessment of telephone receivers, their ability to transfer bacterial infections and the isolation of these microorganisms from receivers, with an emphasis on Staphylococcus aureus and its antibiotic susceptibility pattern. METHODS: 1,591 isolates were obtained from roadside telephone booths in 16 different locations in the Lagos metropolis. They were all characterised using the Cowan and Steel's manual, after which the antimicrobial susceptibility pattern of the most frequently-occurring isolate was determined. RESULTS: Of all the locations, Oshodi was found to have the largest number of microorganisms, with Akoka having the least. The organisms were Providencia, Klebsiella, Citrobacter, Enterobacter, Proteus, Escherichia, Staphylococcus, Bacillus, Streptococcus, Micrococcus and yeast. Staphylococcus spp. was the most frequently occurring, making up 20.2 percent of the total number of isolated organisms, followed by Bacillus (18 percent), while Enterobacter and Citrobacter were the least common. 44 percent of the total Staphylococcus screened for an antimicrobial susceptibility pattern showed high resistance to most of the antibiotics used. CONCLUSION: This result could be related to the emergence of Staphylococcus-resistant strains, especially in a highly-populated area where there are health and hygiene problems and where drugs are abused. Precautions should be taken to prevent the spread of infectious diseases through the use of public telephones.

Campylobacter enteritis in Ilorin, Nigeria.

BACKGROUND: Campylobacterje junilcoli is one of the most commonly identified bacterial causes of acute gastroenteritis worldwide. In Nigeria, it has been reported in the northern part of the country, extensive work has also been done in the south-west leaving behind the middle-belt of the country. OBJECTIVE: To determine the role of Campylobacter jejunilcoli as an agent of diarrhoea in the middle-belt of Nigeria. DESIGN: A prospective case control study. SETTING: University of Ilorin Teaching Hospital (UITH), private hospitals and primary health centers all in Ilorin, Nigeria. SUBJECTS: Three hundred and six children with diarrhoea and 100 without diarrhoea were randomly selected. MAIN OUTCOME MEASURES: Isolation of Campylobacter jejuni/coli from stool samples. Biological characteristics of the isolates. RESULTS: Twenty five (8.2%) of the patients and none of the controls had the organism. Campylobacter jejuni and C. coli were 56% and 44% respectively. All the isolates were susceptible to erythromycin and ciprofloxacin. None of the isolates harboured plasmids. CONCLUSION: C. jejunil coli is an important diarrhoea agent in our geographic environment, further characterisation is desired of these local strains.

Occurrence of Campylobacter species in poultry forms in Lagos area of Nigeria.

Samples of poultry droppings were collected from fifty poultry farms in Lagos area of Nigeria. Campylobacter species were isolated from serially diluted samples using Bolton selective enrichment medium under microaerophilic condition at 42 degrees C. Samples of water, both treated and untreated used in the farms were also analysed in the same manner. Fifteen Campylobacter strains were isolated from the droppings. All the strains were identified using biochemical characteristics differentiating Campylobacter species. They were identified as hippurate positive (C. jejuni) and negative species (C. coli). They were biotype' using the extended scheme described by Lior (1984). Campylobacter coli biotype 'I was the most prevalent (66.66%) followed by C. jejuni biotype 1 (20.00%). The biotype ii of the two species accounted for 6.67% each. The C. jejuni biotype III and IV were not isolated during the study. The preponderance of C. coli indicates that poultry can be a major reservoir for the spread of human campylobacteriosis. Poultry farmers, therefore, must ensure prompt and adequate means of disposal of waste materials as well as maintaining good sanitary conditions.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus in Ilorin, Nigeria.

BACKGROUND: Nosocomial infections caused by methicillin-resistant strains of Staphylococcus aureus constitute significant epidemiologic problems. Defining an outbreak requires the use of rapid and highly discriminatory epidemiologic methods to determine the epidemic strains involved in such outbreak. STUDY DESIGN: A descriptive laboratory based surveillance study for MRSA was undertaken. One hundred and forty seven Staphylococcus aureus isolates from clinical specimens were screened for methicillin resistance at the University of Ilorin Teaching Hospital between January and December 2001. Fifty one (34.7%) methicillin resistant strains recovered were epidemiologically characterized using Eco R1 restriction enzyme analysis of their plasmid DNAs. RESULT: Forty five (88.2%) MRSA isolates were associated with infections and 6 (11.8 %) were colonizing strains; 36 (70.6%) and 15 (29.4%) were hospital and community acquired respectively. Skin and soft tissues were sites of infection in 36 (70.6 %) cases and surgical, emergency and ICU accounted for 33 (64.7%) isolates. All isolates were resistant to more than two antibiotics but sensitive to vancomycin. Forty two (82.4 %) isolates contained plasmids including 9 (21.4 %) that contained more than one plasmid. Restriction Enzyme Analysis of the Plasmid DNA (REAP) divided the isolates into 9 Eco R1 profiles, with profile 2 accounting for 41.7% of all nosocomial infections in the wards, implying that it is endemic. The remaining nosocomial profiles occurred less frequently, suggesting that they are sporadic strains originating from outside the hospital. The community strains showed diverse digestion pattern indicating that they are from different clones. CONCLUSION: The spread of MRSA can be controlled through reinforcement of appropriate use of antibiotics, hand washing and laboratory surveillance for MRSA, particularly in the surgical wards and intensive care units, in order to identify sources of outbreaks.

Multidrug resistance in Salmonella enterica serovar typhi isolated from patients with typhoid fever complications in Lagos, Nigeria.

OBJECTIVE: The aim of this study was to determine the prevalence of Salmonella enterica serovar typhi in patients with pyrexia of unknown origin (PUO), and antimicrobial resistance in strains isolated from patients with S. typhi in Lagos, Nigeria. STUDY DESIGN: In total, 103 patients were included in this study, subdivided into two categories. Category A comprised 74 patients presenting with PUO, and Category B comprised 29 patients harbouring S. typhi who did not respond to initial treatment. METHODS: Blood samples were cultured for bacterial isolates and identified by standard procedures. Susceptibility testing was performed according to the National Committee for Clinical Laboratory Standards. Plasmid DNA extraction was performed using the alkaline lysis method with Escherichia coli v517 used as the standard. Conjugation and transformation experiments were performed using standard methods. For the latter, E. coli K12 HB 101 (ara-14, galK2, hsd 520, lacyl, leu, mtl-1, Pro A2, rec A13, rps L20, sup E44, thii xyl-5) was used as the recipient and plasmid PBR 322 was used as the positive control. RESULTS: The prevalence of S. typhi in PUO was 16.2% (12/74). In all, 25 of 41 (61.0%) cases were multidrug resistant (MDR) with phenotypic-resistant patterns: ACoCT, ACTCeS, ACTCoCe and ACTS were commonly encountered among the strains. Interestingly, four of the strains harbouring similar plasmid DNA were isolated from patients with hepatosplenomegaly, and a strain harbouring a large transferable plasmid of 81.2MDa was isolated from a psychosis patient. CONCLUSIONS: Our study confirmed the circulation of MDR S. typhi in Lagos, Nigeria. Periodic review of antibiotics used in hospitals is essential as the efficacies of chloramphenicol, ampicillin and cotrimoxazole are now doubtful. Ciprofloxacin and ofloxacin are effective drugs for treatment of typhoid fever, but with current trends of drug abuse in Nigeria, resistance is likely to develop. These results, therefore, provide an early warning signal for the prudent use of fluoroquinolone antimicrobials to preserve their usefulness.

Substrate profile variation and drug resistance patterns of â - lactamase producing Shigella species isolated from diarrhoeal patients in Lagos, Nigeria.

The number and trend of antibiotic resistance by Shigella species recovered from food and diarrhoeal stools are on the increase in Nigeria and has resulted in a high frequency of hospitalisation. Increased cost of disease management, and higher mortality in children. This study exposes 51 â-lactamase producing Shigella isolates from Lagos to some newly introduced drugs in the country. The drugs include â-lactam - â-actamase inhibitor antibiotics. â-lctam substrate hydrolysis and inhibitory effects of clavulanate were also investigated in-vitro. Results obtained revealed that all the isolates showed high level resistance to tetracycline, ampicillin, streptomycin, co-trimoxazole and amoxicillin with an MIC range of 128 - 1024 microg/ml. The isolates were susceptible to piperacillin, tobramycin, aztreonam and ofloxacin (0.03 - 8 microg/ ml). 18.2 - 40.9% of S. flexneri and S. dysenteriae showed low level resistance to cefuroxime and cefotaxime (MIC = 4 - 16 microg/ ml). Among the â-lactam - â- lctamase inhibitors tested, only piperacillin-tazobactam showed 100% resistance. Hydrolysis of â -lctam substrate was found to be species dependent in decreasing order of S. flexneri, S. dysenteriae, S. sonnei and S.boydii. An IC50 range of 0.8-2.4 mM was also observed in these isolates. Our data indicate that the incidence of multidrug resistance is high among â-lactamase producing Shigella isolates in Lagos, Nigeria. While the third generation cephalosporins should be used with cautions, some of the newly introduced drugs have the prospects of being used in the future control and management of shigellosis in the country.

Comparison of three PCR methods for detection of Helicobacter pylori DNA and detection of cagA gene in gastric biopsy specimens.

AIM: To comparatively evaluate PCR and other diagnostic methods (the rapid urease test and / or culture) in order to determine which of the three PCR methods (ureA, glmM and 26-kDa, SSA gene) was most appropriate in the diagnosis of Helicobacter pylori (H pylori ) infection and also to evaluate the detection of a putative virulence marker of H pylori, the cagA gene, by PCR in biopsy specimens. METHODS: One hundred and eighty-nine biopsy specimens were collected from 63 patients (three biopsies each) undergoing upper gastroduodenal endoscopy for various dyspeptic symptoms. The PCR methods used to detect H pylori DNA directly from biopsies were the glmM, 26-kDa, ureA and then cagA was used to compare the culture technique and CLO for urease with the culture technique being used as the gold standard. RESULTS: Thirty-five percent of the biopsies were positive for H pylori DNA using the 3 PCR methods, while 68% of these were positive for the cagA gene. Twenty-four percent of the biopsies were negative for H pylori DNA in all PCR methods screened. The remaining 41% were either positive for ureA gene only, glmM only, 26-kDa only, or ureA + glmM, ureA + 26-kDa, glmM + 26-kDa. Out of the 35% positive biopsies, 41% and 82% were positive by culture and CLO respectively, while all negative biopsies were also negative by culture and cagA. Cag A+ infection was also predominantly found in H pylori DNA of the biopsies irrespective of the clinical diagnosis. CONCLUSION: This method is useful for correctly identifying infections caused by H pylori and can be easily applied in our laboratory for diagnostic purposes.