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ABSTRACT: Risk of Campylobacter infection in humans has been associated with many sources, including dogs. C. upsaliensis is the most common species found in canines, and has been occasionally isolated from symptomatic humans. This study aimed to investigate the genetic diversity of 41 C. upsaliensis isolates carried by dogs and from nine isolates carried by humans using Multilocus sequence typing (MLST). We identified considerable genetic diversity amongst the C. upsaliensis isolates from both dogs and humans, identifying 45 different sequence types (STs). All STs were new, apart from that of the reference strain. Only three STs were found in more than one isolate: ST-72 (2 isolates), ST-98 (2 isolates) and ST-104 (3 isolates). ST-104 was the only ST to be encountered in both dogs and humans. Thirty-one of the 45 STs were assigned to one of 13 clonal complexes (CCs). Four of these CCs contained STs originating from both humans and dogs. None of the CCs contained exclusively human isolates, and two isolates from dogs within the same kennel belonged to the same CC. The large amount of diversity found in both dog and human isolates of C. upsaliensis, combined with the relatively small database, made it difficult to assign strains to sources of infection. This emphasizes the need to increase the size of the database. Dog and human isolates occasionally grouped together, however there were insufficient human-derived isolates to determine whether or not dogs are a common source of infection. Although C. upsaliensis infection is rare in humans, dogs still remain a potential source, and are therefore a possible zoonotic risk. Further work is needed to investigate the epidemiology of C. upsaliensis infection in humans.
Veterinary Microbiology 01/2012; 157(3-4):391-7. · 3.33 Impact Factor
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ABSTRACT: Aims: To investigate the prevalence and temporal patterns of antimicrobial resistance in wild rodents with no apparent exposure to antimicrobials.Methods and Results: Two sympatric populations of bank voles and wood mice were trapped and individually monitored over a 2- year period for faecal carriage of antimicrobial-resistant Escherichia coli. High prevalences of ampicillin-, chloramphenicol-, tetracycline- and trimethoprim-resistant E. coli were observed. A markedly higher prevalence of antimicrobial-resistant E. coli was found in wood mice than in bank voles, with the prevalence in both increasing over time. Superimposed on this trend was a seasonal cycle with a peak prevalence of resistant E. coli in mice in early- to mid-summer and in voles in late summer and early autumn.Conclusions: These sympatric rodent species had no obvious contact with antimicrobials, and the difference in resistance profiles between rodent species and seasons suggests that factors present in their environment are unlikely to be drivers of such resistance.Significance and Impact of the Study: These findings suggest that rodents may represent a reservoir of antimicrobial-resistant bacteria, transmissible to livestock and man. Furthermore, such findings have implications for human and veterinary medicine regarding antimicrobial usage and subsequent selection of antimicrobial-resistant organisms.
Journal of Applied Microbiology 02/2011; 110(4):962 - 970. · 2.34 Impact Factor
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ABSTRACT: Rheumatic heart disease (RHD) is an important contributor to cardiovascular disease in children and adults in Yemen. This is the first report to determine the prevalence of RHD among school-children in the city of Aden.
A cross-sectional case-finding survey of RHD was conducted in 6000 school-children aged 5-16 years. Echocardiography was undertaken in those with clinical signs of organic heart disease.
The prevalence of RHD was 36·5/1000 school-children, which is one of the highest reported among school echocardiography surveys in the world. RHD was more common in 10-16-year-old students. RHD was diagnosed in more than one member of the families of 53 (24·2%) of the children. Mitral regurgitation (MR) was detected in 49·8%, 26·6% had MR with mitral valve prolapse and 17·8% had combined MR and aortic regurgitation. Fifty-eight children were diagnosed with congenital heart disease (CHD), representing a prevalence of 9·7/1000. The main types of CHD were mitral valve prolapse, patent ductus arteriosus, atrial septal defect, pulmonary stenosis and aortic stenosis. Congenital mitral valve prolapse found in 36 children was three times more common in males than females. Children with RHD were more likely to be from low-income families with poor housing and greater overcrowding (49·3%, 39·3% and 64·8%) than children with CHD (44·8%, 32·8% and 48·3%, respectively).
The high prevalence of RHD is a major public health problem in Yemen. Urgent screening surveys and an RHD prophylactic programme of appropriate management of group A β-haemolytic streptococcal pharyngotonsilitis are required.
Annals of Tropical Paediatrics International Child Health 01/2011; 31(1):37-46. · 0.90 Impact Factor
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E. D. Carrol,
A. Payton,
D. Payne,
F. Miyajima,
M. Chaponda,
L. A. Mankhambo,
D. L. Banda,
E. M. Molyneux,
H. Cox,
G. Jacobson,
D. F. Carr,
M. E. Molyneux,
J. P. Stewart,
J. P. Quinn, C. A. Hart,
W. E. Ollier
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ABSTRACT: IL-1R antagonist (IL-1Ra) is required for adequate host defense in invasive pneumococcal disease (IPD). The minor allele of an IL1RN gene (C/T) promoter polymorphism (rs4251961) has been shown to be associated with decreased IL-1Ra production in healthy adults. We genotyped 299 children with IPD, and examined 19 IL1RN haplotype-tagging single-nucleotide polymorphisms. Human embryonic kidney HEK293(T) cells were transfected with the promoter reporter plasmid pGL3p containing either allelic variant C (pGL3pCC) or T (pGL3pTT) with or without cotransfection with an expression construct overexpressing the globin transcription factor GATA-1. Plasma IL-1Ra concentrations were significantly higher in nonsurvivors compared with survivors (p < 0.0005), and the C allele of rs4251961 was associated with a significant increase in plasma IL-1Ra concentrations (p = 0.01) during the acute illness of IPD. These findings were validated in a cohort of 276 treatment-naive HIV-infected adults, with borderline significance (p = 0.058). Functional analyses demonstrated that the activity of the promoter constructs containing the T allele increased ~6-fold as compared with basal activity, and that containing the C allele by ~9-fold (p < 0.001) in the presence of GATA-1. Our findings suggest that the IL-1Ra single-nucleotide polymorphism rs4251961 plays a key role in the pathophysiology of IPD and in other human infections.
J Immunol. 01/2011; 186(4):2329-35.
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ABSTRACT: Aims: To investigate the prevalence and temporal patterns of antimicrobial resistance in wild rodents with no apparent exposure to antimicrobials. Methods and Results: Two sympatric populations of bank voles and wood mice were trapped and individually monitored over a 2- year period for faecal carriage of antimicrobial-resistant Escherichia coli. High prevalences of ampicillin-, chloramphenicol-, tetracycline- and trimethoprim-resistant E. coli were observed. A markedly higher prevalence of antimicrobial-resistant E. coli was found in wood mice than in bank voles, with the prevalence in both increasing over time. Superimposed on this trend was a seasonal cycle with a peak prevalence of resistant E. coli in mice in early- to mid-summer and in voles in late summer and early autumn. Conclusions: These sympatric rodent species had no obvious contact with antimicrobials, and the difference in resistance profiles between rodent species and seasons suggests that factors present in their environment are unlikely to be drivers of such resistance. Significance and Impact of the Study: These findings suggest that rodents may represent a reservoir of antimicrobial-resistant bacteria, transmissible to livestock and man. Furthermore, such findings have implications for human and veterinary medicine regarding antimicrobial usage and subsequent selection of antimicrobial-resistant organisms.
Journal of Applied Microbiology 01/2011; · 2.34 Impact Factor
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ABSTRACT: The aims of this study were, firstly, to compare five published methods for the isolation of Arcobacter spp. from animal feces in order to determine the most sensitive and specific method. Second, we analyzed the resulting isolates by multilocus sequence typing (MLST) in order to investigate the diversity of the isolates recovered. Third, we investigated the ability to recover Arcobacter spp. from frozen fecal samples. Seventy-seven fecal samples from cattle, sheep, and badgers were subjected to five isolation methods, based on published methods for the isolation of Arcobacter and Campylobacter spp. Thirty-nine Arcobacter butzleri isolates were analyzed using a multilocus sequence typing scheme. The survival of Arcobacter spp. in frozen samples was investigated by freezing the fecal samples at -80°C for 7 days and then applying the same five isolation methods. The most sensitive and specific method used an Arcobacter-specific broth in conjunction with modified charcoal cefoperazone deoxycholate agar (mCCDA) with added antibiotics. Freezing of fecal samples led to a reduction in the recovery of Arcobacter spp. by approximately 50%. The 39 allelic profiles obtained by MLST could be divided into 11 sequence types (STs). We have identified the most sensitive and specific method for the isolation of Arcobacter spp. from animal feces and demonstrated that the freezing of fecal samples prior to isolation reduces arcobacter recovery. MLST analysis of the isolates revealed a high level of diversity.
Applied and environmental microbiology 12/2010; 77(5):1646-50. · 3.69 Impact Factor
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ABSTRACT: Dog ownership is considered a risk factor for campylobacteriosis in humans. This study investigated the prevalence and shedding of Campylobacter spp. in kennelled dogs. Faecal samples (n=399) were collected in longitudinal studies from 52 dogs in two kennels. Campylobacter spp. were isolated using charcoal-based selective agars and direct PCR. The prevalence of Campylobacter spp. in dogs in boarding kennels ranged from 46% (95% CI 22, 72) on entry, to 50% (95% CI 30, 70) overall, and in dogs in 'rescue' kennels from 68% (95% CI 49, 84) on entry to 73% (95%, CI 56, 87) overall. C. upsaliensis was isolated from 62% (95% CI 48, 73) of the dogs, whilst C. jejuni was isolated from 15% (95% CI 7, 26) of animals. The majority of infected dogs entered the kennels already carrying Campylobacter spp., and remained infected throughout their stay. However, in some cases, shedding appeared to commence after kennelling. Given that the prevalence of C. upsaliensis and C. jejuni was relatively high in dogs from both boarding and rescue kennels, such animals may pose a zoonotic risk.
The Veterinary Journal 11/2010; 190(2):249-54. · 2.24 Impact Factor
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ABSTRACT: A survey was undertaken of the etiology of acute gastroenteritis in children <16 years of age in Antananarivo, Madagascar, from May 2004 through May 2005. With use of electron microscopy of fecal specimens, 104 (36%) of 285 children were found to be infected with rotavirus. Rotavirus strain characterization was undertaken using enzyme-linked immunosorbent assay, electropherotyping, reverse-transcription polymerase chain reaction genotyping, and nucleotide sequencing. The predominant group A rotavirus strain types identified were P[4]G2 (62%) and P[8]G9 (23%). Nucleotide sequence analysis of the VP7 genes of selected Malagasy G2 and G9 strains demonstrated similarity with those of other recently identified African rotavirus strains belonging to the same genotype.
The Journal of Infectious Diseases 09/2010; 202 Suppl:S175-9. · 6.41 Impact Factor
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ABSTRACT: A total of 969 isolates of Campylobacter jejuni originating in the Preston, Lancashire postcode district over a 3-year period were characterized using multi-locus sequence typing. Recently developed statistical methods and a genetic model were used to investigate temporal, spatial, spatio-temporal and genetic variation in human C. jejuni infections. The analysis of the data showed statistically significant seasonal variation, spatial clustering, small-scale spatio-temporal clustering and spatio-temporal interaction in the overall pattern of incidence, and spatial segregation in cases classified according to their most likely species-of-origin.
Epidemiology and Infection 03/2010; 138(10):1384-90. · 2.84 Impact Factor
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ABSTRACT: The role of coronaviruses in paediatric gastro-enteritis is not well defined. We investigated the detection rate and epidemiological features of infection with coronavirus in children receiving hospital care for acute gastro-enteritis in Maddina, Saudi Arabia.
Stool specimens were collected from children less than 5 years of age who were either hospitalised in Maddina or given oral rehydration therapy as outpatients between April 2004 and April 2005. Coronaviruses were detected by electron microscopy.
Coronaviruses were detected in 63 (6%) of 984 children with acute gastro-enteritis and were more commonly detected in outpatients (47/423, 11%) than in inpatients (16/561, 3%). The median age (range) of children with coronavirus infection was 42 months (10-60). Coronaviruses were detected throughout the year with the highest detection rate at the end of the winter season.
Coronaviruses were commonly identified in children with diarrhoea in Saudi Arabia. Their role in paediatric gastro-enteritis warrants further evaluation.
Annals of Tropical Paediatrics International Child Health 01/2010; 30(1):45-50. · 0.90 Impact Factor
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ABSTRACT: Samples of faeces were taken from 183 healthy pet dogs in a census-based, cross-sectional study in Cheshire; culture methods were used to detect any Campylobacter species and a direct PCR was used to detect Campylobacter upsaliensis. Forty-six of the dogs were positive for C upsaliensis by either culture or direct PCR, giving a prevalence of 25.1 per cent (95 per cent confidence interval [CI] 19.0 to 32.1 per cent). One sample was positive by culture for Campylobacter jejuni (95 per cent CI 0.0 to 3.0 per cent) and one for Campylobacter lari. Multivariable logistic regression identified risk factors for the carriage of C upsaliensis by a dog as: living with another dog that also carried C upsaliensis; being small rather than medium-sized; being less than three years old; living in a household that kept fish; being fed commercial dog treats; and being fed human food titbits, particularly in the dog's bowl.
The Veterinary record 10/2009; 165(18):526-30. · 1.25 Impact Factor
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B N Parsons,
A J Cody,
C J Porter,
J H Stavisky,
J L Smith,
N J Williams,
A J H Leatherbarrow, C A Hart,
R M Gaskell,
K E Dingle,
S Dawson
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ABSTRACT: Campylobacter is a major cause of human gastroenteritis worldwide. Risk of Campylobacter infection in humans has been associated with many sources, including dogs. This study aimed to investigate whether C. jejuni carried by dogs could potentially be a zoonotic risk for humans and if there were common sources of C. jejuni infection for both humans and dogs. Multilocus sequence typing (MLST) together with macrorestriction analysis of genomic DNA using SmaI and pulsed-field gel electrophoresis (PFGE) were both used to analyze 33 C. jejuni isolates obtained from various dog populations, including those visiting veterinary practices and from different types of kennels. MLST data suggested that there was a large amount of genetic diversity between dog isolates and that the majority of sequence types found in isolates from these dogs were the same as those found in isolates from humans. The main exception was ST-2772, which was isolated from four samples and could not be assigned to a clonal complex. The most commonly identified clonal complex was ST-45 (11 isolates), followed by ST-21 (4 isolates), ST-508 (4 isolates), and ST-403 (3 isolates). The profiles obtained by macrorestriction PFGE were largely in concordance with the MLST results, with a similar amount of genetic diversity found. The diversity of sequence types found within dogs suggests they are exposed to various sources of C. jejuni infection. The similarity of these sequence types to C. jejuni isolates from humans suggests there may be common sources of infection for both dogs and humans. Although only a small number of household dogs may carry C. jejuni, infected dogs should still be considered a potential zoonotic risk to humans, particularly if the dogs originate from kennelled or hunt kennel dog populations, where the prevalence may be higher.
Journal of clinical microbiology 09/2009; 47(11):3466-71. · 4.16 Impact Factor
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ABSTRACT: The concept of colonisation resistance is based on animal experiments. This study was designed to quantify the defence against colonisation by ingested exogenous microorganisms in human volunteers, and to measure the effect of oral amoxycillin on colonisation resistance. Ten healthy volunteers were challenged with increasing concentrations of a test strain of Klebsiella oxytoca, 103, 105, 107and 109 colony forming units (cfu). Oral washings and faecal specimens were collected three times a week and were processed qualitatively and quantitatively. After a challenge dose of 107 cfu transient intestinal colonisation occurred in one volunteer, and after 109 cfu in two volunteers. The experiment was repeated after three days of amoxycillin intake with a challenge dose of 105 cfu. In five of nine volunteers the exogenous microorganisms were able to colonise the intestine, with severe clinical symptoms of profuse diarrhoea in one volunteer. This study confirms the existence of colonisation resistance in humans and the adverse effect of amoxycillin.
07/2009; 1(3):169-177.
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B N Parsons,
C J Porter,
R Ryvar,
J Stavisky,
N J Williams,
G L Pinchbeck,
R J Birtles,
R M Christley,
A J German,
A D Radford, C A Hart,
R M Gaskell,
S Dawson
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ABSTRACT: Campylobacteriosis is a major cause of gastroenteritis in humans and some studies have suggested that dog ownership is a risk factor for the condition. To determine the prevalence, species distribution, and risk indicators for Campylobacter spp. infecting dogs attending veterinary practices in UK, faecal samples were collected in a cross-sectional study from 249 dogs with and without clinical signs. The prevalence of Campylobacter spp. was 38% (95% CI 32, 44), with Campylobacter upsaliensis accounting for 94 (98%) of the isolates and Campylobacter jejuni for the remainder. Multivariable analysis indicated that younger dogs were more likely to carry C. upsaliensis and the high prevalence of this pathogen supports the hypothesis that dogs, particularly younger animals, may be an important source of C. upsaliensis infection for humans. However the prevalence of C. jejuni, the most common Campylobacter spp. associated with disease in humans, was low (1.2%, 95% CI 0.3, 3).
The Veterinary Journal 03/2009; 184(1):66-70. · 2.24 Impact Factor
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ABSTRACT: Acute respiratory infections (ARI) cause significant childhood mortality. Nutritional homeostasis, particularly micronutrient levels, is important in modulating response to infection. More information is required regarding micronutrient levels in ARI viral infections, especially newly identified viruses such as human metapneumovirus (HMPV).
To describe zinc, copper, selenium and vitamins A and E concentrations in children with respiratory syncytial virus (RSV) and/or HMPV in relation to levels of C-reactive protein (CRP).
The presence of RSV/HMPV in nasopharyngeal aspirates (NPA) was identified in 246 children using RTPCR. Zinc, copper, selenium and vitamins A and E concentrations were measured using inductive coupled plasma mass spectrometry and high performance liquid chromatography.
183 children had RSV, 39 had HMPV and 24 were co-infected. Zinc concentrations were lower in children with HMPV than in children with RSV or RSV/HMPV co-infection. Copper concentrations were lower in children with RSV than in children with RSV/HMPV or HMPV and zinc/copper ratios were lower in children with HMPV/RSV or RSV than in children with HMPV alone. Retinol and a alpha-tocopherol were lower in children with RSV than in children with HMPV. Most children had low selenium concentrations. Children with RSV and raised CRP (>5 mg/L) had higher copper and lower zinc/copper ratios than those with low CRP (< or =5 mg/L). Children with HMPV and raised CRP had higher copper and lower zinc concentrations than children with low CRP. Children with RSV/HMPV and raised CRP had higher copper concentrations. Children with RSV/HMPV and raised CRP had higher a alpha-tocopherol concentrations.
The profiles of micronutrients differ in children with RSV and HMPV and are confounded by CRP. These results may guide strategies for micronutrient supplementation in ARI.
Annals of Tropical Paediatrics International Child Health 03/2009; 29(1):35-40. · 0.90 Impact Factor
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ABSTRACT: Cattle faeces are considered the most important reservoir for human infection with Escherichia coli O157. We have previously described shedding of E. coli O157 in the faeces of naturally infected cattle cohorts. However, the data require further investigation to quantify the uncertainty and variability in the estimates previously presented. This paper proposes a method for analysing both the presence and the quantity of E. coli O157 in cattle faecal samples, using two isolation procedures, one of which enumerates E. coli O157. The combination of these two measurements, which are fundamentally different in nature and yet measuring a common outcome, has necessitated the development of a novel statistical model for ascertaining the contribution of the various components of variation (both natural and observation induced) and for judging the influence of explanatory variables. Most of the variation within the sampling hierarchy was attributable to multiple samples from the same animal. The contribution of laboratory-level variation was found to be low. After adjusting for fixed and random effects, short periods of increased intensity of shedding were identified in individual animals. We conclude that within-animal variation is greater than between animals over time, and studies aiming to elucidate the dynamics of shedding should focus resources, sampling more within than between animals. These findings have implications for the identification of persistent high shedders and for assessing their role in the epidemiology of E. coli O157 in cattle populations. The development of this non-standard statistical model may have many applications to other microbial count data.
Journal of The Royal Society Interface 02/2009; 6(31):169-77. · 4.40 Impact Factor
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ABSTRACT: A 13-month study of children presenting with acute diarrhoeal disease at hospitals and rehydration clinics in Antananarivo, Madagascar, was undertaken between May 2004 and May 2005. Cryptosporidiosis accounted for diarrhoea in 12 (5.6%) of the 215 children investigated. Cases of cryptosporidiosis were detected only in the rainy season, and the median age of cases was 13.5 months (range=1 day-27 months). As 11 of the cases of cryptosporidiosis were caused by Cryptosporidium hominis and only one by C. parvum, most of the cases were probably the result of anthroponotic transmission. GP60/45/15 gene polymorphisms indicated that the causative pathogens were of subtypes Ia, Id, Ie and IIc.
Annals of Tropical Medicine and Parasitology 07/2008; 102(4):309-15. · 1.43 Impact Factor
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ABSTRACT: Bacteroides thetaiotaomicron NCTC 10582 grown in media containing pig gastric mucin was found to be capable of producing all the glycosidases required to degrade the carbohydrate moieties of human colonic mucin. These are α-fucosidase, β-galactosidase, α-N-acetylgalactosaminidase, β-N-acetylglucosaminidase and neuraminidase. Moreover, a novel glycosulphatase was identified using glucose-6-sulphate as substrate. This enzyme has a Km of 43·4 mmol/l and a pH optimum of 5·0. The bacteria, when cultured for 24 h in broth, were capable of removing 18% of [35S]-sulphate from [35S]-labelled mucin and of removing 15% of [3H]-glucosamine from [3H]-glucosamine-labelled human colonic mucin. The results suggest that this bacterium is likely to play an important role in mucus degradation in the human colon.
Letters in Applied Microbiology 06/2008; 13(2):97 - 101. · 1.62 Impact Factor
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ABSTRACT: Mucosal biopsies from canine jejunum and ileum were established in organ culture, and remained structurally intact for up to 72 hours. Infection of such mucosal biopsies with an enteropathogenic Escherichia coli (EPEC) 0111 results in the characteristic loss of brush border observed during natural and induced infection in other animal species. Within four hours of infection, bacteria had become attached to the mucous overlying the microvilli and by eight hours had penetrated between the microvilli resulting in partial loss of the brush border. After 24 hours incubation there was almost complete effacement of the brush border with cup and pedestal formation at sites of bacterial adherence. We conclude that EPEC should be considered as a potential cause of diarrhoea in the dog and that organ culture of canine mucosal biopsies provides a useful model for studying infection with EPECs and other enteric pathogens.
Journal of Small Animal Practice 06/2008; 31(12):591 - 594. · 1.00 Impact Factor
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Journal of Applied Microbiology 03/2008; 74(S22):33S - 42S. · 2.34 Impact Factor
