antibiotic susceptibilities of salmonella species ... agglutination test kit. ... antibiotic...

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Mzunguetal.ArchMedBiomedRes.2016;3:39-51.doi:10.4314/ambr.v3i1.6

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Antibiotic susceptibilities ofSalmonella species prevalent amongchildren of 0-5 yearswith diarrhea inKatsinastate,NigeriaIgnatius Mzungu1, Helen I Inabo2, Stephen O Olonitola2,MaryamAminu2ABSTRACTThis study was conducted to assess the prevalence of Salmonellaspecies among children having diarrhea in Katsina State, Nigeria. Atotal of 220 diarrhea stool samples of children aged five years andbelow(0-5years)werecollectedandscreened forSalmonella speciesusing culture technique. Presumptively positive isolates were furtherscreened biochemically and serologically, using MicrogenTMEnterobactericeaeIDkitandMicrogenTMSalmonellarapidconfirmatorylatex agglutination test kit. Antibiotic susceptibility of confirmedisolates was carried out and resistance patterns of the isolatesdetermined. Thehighest incidencewasobserved in childrenof 13-24monthsofageandthe least inchildrenof0-12monthsofage.Therewasahigherprevalence inmalethanfemalechildren.All the isolatesscreened were resistant to Ampicillin and Amoxicillin and of these,90.9% were resistant to Amoxicillin-clavulanic acid and 45.5% wereresistant to Nalidixic acid. However, 100% were sensitive toGentamicin,90.9%weresensitivetoCiprofloxacinandCefotaximeand9.09% showed reduced susceptibility to Ciprofloxacin. Both theoccurrence of Salmonella in children and their resistance tomultipleantibioticsasobservedareofpublichealthsignificance.Theveinofthisstudy underscores the importance of routine monitoring of theincidenceofSalmonellaandcontinuedhealtheducationofcaregivers.KEY WORDS: Antibiotic resistance; Prevalence of Salmonella;Childhood diarrhea; Katsina State; Antibiotics susceptibility ofSalmonellaINTRODUCTIONDiarrhea is defined as the passage of three or more loose orliquid stoolsperday (ormore frequentpassage than isnormalfortheindividual).Itisusuallyasymptomofaninfectionintheintestinal tract, which can be caused by a variety of bacterial,viral and parasitic organisms. Infection is spread throughcontaminatedfoodordrinking-water,orfromperson-to-personasaresultofpoorhygiene.Diarrhealdiseasescausedbyentericinfectionsremainaleadingglobalhealthproblem.Twotofourbillionepisodesofinfectiousdiarrhea have been estimated to occur annually in developing

To cite: Mzungu I, Inabo HI,Olonitola SO, Aminu M.Antibiotic susceptibilities ofSalmonella species prevalentamong children of 0-5 yearswith diarrhea in Katsina state,Nigeria.ArchMedBiomedRes.2016;3(1):39-51. doi:10.4314/ambr.v3i1.6PublicationhistoryReceived:November20,2015Revised:March20,2016Accepted:March21,2016OpenAccessThis is an Open Access articledistributed inaccordancewiththe Creative CommonsAttribution Non Commercial(CC BY-NC 3.0) license, whichpermits others to distribute,remix, adapt, build upon thiswork non-commercially, andlicense their derivative workson different terms, providedthe original work is properlycited and the use is non-commercial.CrossRefLinkhttp://dx.doi.org/10.4314/ambr.v3i1.6CorrespondencetoIgnatiusMzungu;[email protected]



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countries,resultingin3to5milliondeaths.1

It is a common cause of infant deaths indevelopingcountries,especiallywheresafedrinkingwaterandadequatesanitationandhygiene is unavailable.2,3 It remains thesecond leading cause of death amongchildren under five globally.4 Diarrhoealdiseasekills2,195childreneveryday(about801,175 each year), accounting for 1 in 9child deaths worldwide.5 As at the end of2015,Nigeriastillrankedsecondamongthetop 15 countries with high child mortalityduetodiarrheaandpneumonia.6The major causes of diarrheal illnessinclude,amongothers, limitedaccess toorpoor quality of water, poor food hygiene,andsanitation.Reportsfromdifferentpartsof the world have implicated variouspathogens including Parasites (Giardialamblia, Entamoeba histolytica), bacteria(Escherichia coli, Salmonella species,Shigella species, Campylobacter jejuni,Klebsiella species, Enterobacter speciese.t.c.) and viruses like the Rotavirus, withtheoutbreakof infantilediarrhealdisease7-12,allofwhichareknowntobeendemic inessentially all developing countries.However in developing countries likeNigeria, infantilediarrheadisease is grosslyunder-reported and the incidence under-estimated, this isattributedtopovertyandignorance among the affected group whoconstitute up to 80% of the population ofthe area13, the lack of coordinatedepidemiological surveillance system,inadequacy of laboratory facilities forculture and unsafe water from all sourcescontributesignificantlytotheglobalburdenofdisease.14Salmonella, a genus of Gram-negative rod-shaped bacteria of the familyEnterobacteriaceae,causesawiderangeofhuman diseases, such as enteric fever,gastroenteritis, endocarditis, andbacteraemia.It isoneofthemostcommonandwidelydistributedfoodbornediseases.It constitutes amajorpublic healthburden

and represents a significant health cost inmany countries. Millions of human casesarereportedworldwideeveryyearandthedisease results in thousands of deaths.15Although infections with non-typhoidalSalmonellae usually cause self-limitingdiarrhealillness,serioussequelae,includingmeningitis, sepsis, and death, may occur,especially among infants and elderlypersons.16In recent years problems related toSalmonella have increased significantly,both in terms of incidence and severity ofcases of human salmonellosis. Socio-demographic factors (age, education,income etc.), environmental and sanitationfactors(pooraccesstoagoodwatersourceand poor sanitation) and climatic factors(rainfall, temperature and humidity) arethought to be related to incidence andspatial distribution of diarrhea.17 Thefrequency and gravity of these infectionsare affected by hygienic conditions,malnutrition, and the excessive use ofantibiotics that select for multi-drugresistantstrains.18In Nigeria, morbidity associated withillnessesduetoSalmonella continues tobeon the increase and, in some cases,resultingindeath.Newconcernshavebeenidentified as since the beginning of the1990s, strains of Salmonella which areresistanttoarangeofantibiotics, includingfirst-choice agents for the treatment ofhumans, such as chloramphenicol and co-trimoxazole, and the third-generationcephalosporins, have emerged and arethreatening to become a serious publichealth problem19 (Table 1). AlthoughFluoroquinolones have been found to beefficacious both in vitro and in vivo in thetreatment of severe Salmonella-associatedillnesses,strainswithreducedsusceptibilitytociprofloxacinamongtravelershavebeenreported in someparts of the globe20. Thisresistance results from the use ofantimicrobials both in humans and animal



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husbandry. Multi-drug resistance to"critically important antimicrobials" iscompoundingtheproblems.Table1:ReviewofoccurrenceofantibioticresistantSalmonellaphenotypesinNigeriaAuthor(year) Location Antibiotic (% of

resistantisolates)Olowe et al.(2007)

Osogbo AML (93.3), SXT(93.3), AMP(86.9),STP(82.6),CIP(30.4)

Ifeanyi et al.(2010)

Abuja AML (55.6), AMC(55.6), CPL (55.6),CFR(55.6)

Sule et al.(2012)

Anyigba PFX (100), CIP(100), AMC (100),GEN (100), SXT(100), AMP (100),CPL (20.0), NAL(20.0), STP (40.0),OFL(10.0)

Ajibade(2013)

Ekiti-State STP (100), CHL(100), CTX (100),AMP (100), NAL(100),GEN (96.0),CIP (69.0), TET(100),SXT(100)

Moses et al.(2014)

Ile-Ife NIT (100), CFZ(97.2), GEN(94.2), AML(52.2), AMC(55.1), CFZ (89.9),SXT(44.9)

Galadima andKolo(2014)

Minna CHL (100), STP(100)

Key: STP-Streptomycin, AML-Amoxycillin, AMC-Amoxycilin-clavulanicacid,CPL-Cephalexin,CFR-Cefuroxime, PFX-pefloxacin, AMP-Ampicillin,NIT-nitrofurantoin, CFZ-ceftriazone, GEN-gentamicin,CHL-chloramphenicol,NAL-Nalidixicacid,CTX-CefotaximeMETHODOLOGYStudyPopulationThestudywashospitalandclinicbased.Thestudy population included infants andyoungchildrenoffiveyearsandbelowwho

reported to the hospitals or clinics withdiarrhea symptoms or gastroenteritis. TheflowchartofthestudyisoutlineinFigure1.

Figure1:FlowchartofMethodology

Keys: SSA - Salmonella/Shigella agar; XLD -XyloseLysineDeoxocholateControlsubjects:Thesewerechildrenofthesame age group, who had no history ofdiarrhea, at least three weeks precedingsampling.Inclusion criteria: Children aged below 5years, presenting with acute diarrheaattending public and private hospitalswithin the study area and who have givenconsent.Exclusion criteria: Includechildrenabove5years,ofbothsexesandlackofconsent.Operationaldefinition:Diarrheacaseinthisstudy was defined as a child passing looseor watery stool or a bloody stool three ormore times in 24-hour period as reported



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by parents. The control subjects werechildren presented for illnesses other thandiarrheaandwithnohistoryofdiarrhea inthe three weeks preceding sampling.Histories were taken from each child frominformedandconsentedparents/caregiversbefore sample collection by the attendingqualified health worker. All the relevantdata (demographic, clinical, and laboratorydata)wererecorded.Ethical Consideration: EthicalapprovalwasobtainedfromtheKatsinaStateMinistryofHealth(RefNo:Perm.Sec./065-35554)andtheethicscommitteeofeachhospitalunderstudy before the commencement of thestudy. In the course of this research,individuals’ anonymity was maintained,good laboratory practice/quality controlwasensuredandevery findingwas treatedwith utmost confidentiality and for thepurposeoftheresearchonly.SampleCollectionAbout5-10gofstoolwasscoopedintowell-labeledsterilewidemouthuniversalsamplebottleusingplasticspoon.Whereitwasnotpossible to obtain stool, rectal swab wasmade using sterile swabs and placed inphysiological saline in properly labeledsample bottles. Sample collection formswere filled foreach sample.This containedinformation such as: color of stool, texture(formed, semi-formed, uniform, fluid) andpresence of blood,mucus, pus in samples.AllsamplesweretransportedinicepackstotheBacteriology laboratory,DepartmentofMicrobiology Ahmadu Bello University,Zariaforanalysis.MicrobiologicalanalysisofstoolsamplesAloopfulof liquidstoolorfecalsuspensionwas enriched in selenite F broth for 24hours at 37°C and then sub-cultured onMacConkey agar (Oxoid) and SalmonellaShigella agar (Oxoid). The plates wereincubated at 37°C for 24 hours. Theresultant isolates were purified for further

tests.NonlactosefermentingorganismsonMacConkey agar (MCA) and SalmonellaShigella agar (SSA) with colorless or palecolonies were purified on Xylose LysinDeoxocholate (Oxoid) agar, and stored onNutrient agar (Oxoid) slants for furtherstudies. Media preparations were carriedout according to manufacturer'sinstructions.BiochemicalcharacterizationofisolatesAll isolates were biochemicallycharacterizedusingtheMicrogenTMGnA+B-ID System for Enterobacteriaceae. Testswere performed strictly following themanufacturer'sinstructions.Serological identification of SalmonellaisolatesSerology tests were performed on allisolates positive for Salmonella bybiochemical characterization usingMicrogenTM Salmonella kit; a rapidconfirmatory latex agglutination test forSalmonella. All tests were performedaccording to the manufacturer'sinstructions.Determination of antibiotic susceptibilitypatternofisolatesIsolatesfrombothwaterandstoolsamples,earlier characterized were tested forsusceptibilitytothefollowing10antibiotics,commonly used in humans for thetreatment of Gram negative pathogens;Gentamicin (10µg), Ampicillin (10µg),Amoxycillin (30µg),Trimethoprim/Sulphamethoxazole (1:19)(25µg), Ciprofloxacin (5µg), Nalidixic acid(30µg), Tetracycline (30µg), Cefotaxime(30µg), Chloramphenicol (30µg),Amoxicillin/clavulanicacid(2:1),(Oxoid).Antimicrobial susceptibility testing wasperformed for all Salmonella isolates usingthediskdiffusionmethodand resultswereinterpretedusingthecriteriaofClinicalandLaboratory Standards Institute guidelines.21



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Briefly,eachstoredisolatewassub-culturedon SS agar (Oxoid). When a pure culturewas obtained, a loopful of bacteria wastakenandtransferredtoatubecontaining2ml 0.85% saline (pH 7.2) andmixed gentlyuntil it formed a homogenous suspension.The turbidity of the suspension was thenadjusted to the optical density of aMcFarland unit of 0.5 to standardize theinoculumsize;0.1mlofthesuspensionwastransferred onto the surface of MuellerHintonagar(Oxoid)andevenlyspreadusinga sterile bent glass rod. The inoculatedplateswereleftatroomtemperaturetodryfor 10-15 minutes. With the aid of diskdispenser,asetofantibioticdiskswasthenplacedgentlyonthesurfaceoftheMuellerHinton agar (3 antibiotic discs per plate).The discs were gently pressed onto thesurface of themediumwith sterile forcepsto ensure firm contact and incubated at37oCfor24hours.Diametersofthezoneofinhibition around the antibiotic discs weremeasuredtothenearestmillimeterusingarule and the isolates were classified as

sensitive (S) or resistant (R), based on CLSIbreak points. Intermediates were reportedasisolateswithreducedsusceptibility.RESULTSA total of 220 stool samples of childrenaged fiveyearsandbelow,presentingwithdiarrhea, were collected and investigatedfor Salmonella infection along with 22control samples from apparently healthychildrenwithin the studyarea. Of the220studysubjects,119(54.1%)weremalesand101(45.9%)werefemales.Prevalence of Salmonella species amongthestudypopulationFigure 2 shows the overall prevalence ofSalmonella species among the studysubjects within the study area. Out of 220samples collected from children withdiarrhea, 11 were positive for Salmonellaspecies, giving the overall prevalence of5.0%inthearea.

Figure2:PrevalenceofSalmonellaspeciesamongthestudypopulation

0

50

100

150

200

250

TotalNo.screened No.Positive Prevalence

220

115.00%

Counts



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PrevalenceofSalmonellaspp.accordingtoageandgenderofthestudypopulationTable2showstheprevalenceofSalmonellaspeciesinrelationtoageandgenderofthestudysubjects.Ahighfrequencyofisolationoccurred among children aged 13-24months and 25-36 months (36.4%), and a0.0% prevalence among 0-12 months old.Male children were more affected thantheir female counterparts. There was astatistically significant difference in theoccurrence of Salmonella with respect toageandgender.Antibiotic susceptibility of SalmonellaisolatesTable 3 shows the result of antibioticsusceptibility of Salmonella isolates wherethe organisms showed 100% resistance tothe β-lactams: Ampicilin, Amoxycillin and90.9% resistance to Amoxicillin-clavulanic

acidand63.6%ofisolateswereresistanttoSulphamethoxazole Trimethoprim. Theisolateswere,however,100%susceptibletotheaminoglycoside-Gentamicin,90.9%and45.5% susceptible to the quinolones(Ciprofloxacin and Nalidixic acid)respectively. A sensitivity of 63.6% toChloramphenicolwasalsoobserved.AntibioticresistancepatternofSalmonellaisolatesTable 4 shows the patterns of antibioticresistance by Salmonella isolates in thestudy. Two isolates showed resistance tothree antibiotics. One isolatewas resistantto 4 antibiotics, 2 were resistant to 5antibiotics, 3 isolates showed resistance to6 antibiotics, and 3 isolates showedresistanceto7antibiotics.Variousantibioticresistance patterns were exhibited by theisolatesasshowninthetable.

Table2:PrevalenceofSalmonellaspp.accordingtoageandgenderofthestudypopulation

Agegroup Male Female Total

No.Examined

No.(%)positive

No.Examined

No.(%)positive

No.Examined

No.(%)positive

0-12 37 0(0.0) 25 0(0.0) 62 0(0.0)

13-24 40 2(18.2) 53 2(18.2) 93 4(36.4)

25-36 15 3(27.3) 21 1(9.0) 36 4(36.4)

37-48 15 1(9.0) 9 1(9.0) 24 2(18.2)

49-60 3 1(9.0) 2 0(0.0) 5 1(9.0)

Total 110 7(63.6) 110 4(36.4) 220 11(100)

χ2=1.454,P≤0.05,df=4



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Table3:AntibioticsusceptibilityofSalmonellaspeciesisolatedfromdiarrheicstools

Antibiotic Susceptibilityofisolatestoantibiotics(n=11)

Susceptible Intermediate Resistant

Ampicilin 0 0 11(100%)

Amoxycillin 0 0 11(100%)

AmoxicillinClavulanicacid

0 1(9.09%) 10(90.9%)

Gentamicin 11(100%) 0 0

SulphamethoxazoleTrimethoprim

4(36.4%) 0 7(63.6%)

Chloramphenicol 7(63.6%) 1(9.09%) 3(27.3%)

Cefotaxime 10(90.9%) 0 1(9.09%)

Tetracycline 2(18.2%) 0 9(81.8%)

Ciprofoxacin 10(90.9%) 1(9.09%) 0

Nalidixicacid 5(45.5%) 4(36.4%) 2(18.2%)

Table4:AntibioticresistancepatternofSalmonellaspeciesisolatedfromdiarrheicstool

samplesNo.ofisolates No.ofantibioticsresistant

toResistancepattern

1 3 AMP,AMC,CTX

1 4 AMP,AMC,TET,AML

2 6 AMP,AMC,CTX,TET,SXT,AML

2 7 AMP,AMC,CTX,TET,SXT,CHL,AML

1 5 AMP,AMC,TET,SXT,AML

1 5 AMP,AMC,CTX,TET,AML

1 6 AMP,AMC,TET,NAL,SXT,AML

1 7 AMP,AMC,TET,NAL,SXT,CHL,AML

1 3 AMP,CTX,AML

Key: AMP- Ampicilin; AML- Amoxycillin; AMC- Amoxicillin Clavulanic acid; SXT-Sulphamethoxazole Trimethoprim; CHL- Chloramphenicol; CTX- Cefotaxime; TET- Tetracycline;NAL-Nalidixicacid



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DISCUSSIONDiarrheal diseases caused by entericinfections remain a leading global healthproblem. It is a common cause of infantdeaths in developing countries, especiallywhere safe drinking water and adequatesanitation and hygiene is unavailable.2,3Identification of the enteropathogensinvolved indiarrhealdisease in thecountryis an essential step towards theimplementationof effectiveprimaryhealthcare activities against the disease.22 In thisstudy, a prevalence of 5.0% was observedfor Salmonella species among 220 childrenwith diarrhea. This low prevalence reflectsthe fact that diarrhea disease in childrenhasmultipleetiologiesrangingfromviruses,parasites and other bacteria agents. ThesehavebeenreportedwithvaryingprevalencearoundtheworldincludingNigeria.Ogbuetal23 reported the occurrence of Rotavirus(23.3%),E. coli (15.4%), Salmonella species(11.3%), Klebsiella species (7.3%) andEnterobacter species (9.6%) as the mostpredominant etiological agents of diarrheachildren in Abakaliki south-east Nigeria.Shigella,Yersiniaenterocolitica,Entamoebahistolytica and Giadia lamblia were alsorecovered in that study. Similarly,Okoloetal reported the occurrence of E. coli,Salmonella Campylobacter and Shigellaspecies among children in Anyigba NorthCentral Nigeria.24 Ifeanyi et al observed E.coli, Salmonella Typhi, Klebsiellapneumoniae,Staphylococcusaureus amongothers25; and Akingbade et al26 isolated E.coli, Salmonella and Shigella. However,KorieetalobservedonlyEnteropathogenicE.colifromtheirreportinEnugu.27Therewasnocaseobservedamong the22control subjects included in the study,indicatinga lowcarrierrateof theagent inchildren of this age (≤5years), which isprobablyduetotheirlowerimmunestatus,and virulence of Salmonella species.However, carrier caseshavebeen reported

among primary school pupils in a study inAkure,Nigeria.28

TheoccurrenceofSalmonellaspeciesinthisstudyisinconformitywiththefindingsfromBissau, Guinea Bissau29, Hong Kong30, SaoPaulo, Brazil31, Abakaliki, South–easternNigeria,23 and Anyigba, North CentralNigeria24.TheprevalenceofSalmonellaspecies(5.0%)inthestudysubjectsagreewiththereportsof similar studies in Lagos32, Bangladesh33,Korea34, and Ghana.35 However, theprevalencerateishigherthanthatobtainedbyIfeanyietal25whoreportedaprevalenceof3.2%fromastudyinAbuja,Nigeria.Thiscouldbeduetothesignificantdifferenceininfrastructuraldevelopment,socioeconomicstatusandeducationallevelofpeoplelivingin the area of study (Abuja). This reporthowever, shows lower Salmonellaprevalence than that reported by Ogbu etal. (11.3%),23 Duru et al36 reported aprevalence of 10.0%, Ike and Damola37reported10.7%,whileGaladimaandOkoloobservedaprevalenceof16.6%.3

The disparity may be attributable todifferences in study design, patients’selection, differing environmentalconditions and behavioral patterns ofpeople in those regions. Occurrence ofdiarrhea has been associated with factorsincluding lack of education ofmother, lackof exclusive breastfeeding, poor nutritionalstatus, immunization status, personalhygiene, overcrowding, garbage disposal,source of water supply, and toilet facility,which are also predominant in the studyarea.38

The result fromthis studyshowed that themajority of Salmonella species weredetected among children of between theages 13-24 months and 25-36 months(36.0%). Abdullahi et al39 observed thehighest prevalence among childrenbetween 20-24 months of age. Similarly,Okolo et al24 reported the prevalenceamong children between ages 13-24



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months (28.1%). These findings could bedue to combinedeffectsofdeclining levelsofmaternally acquired antibodies, the lackof active immunity in the infant, theintroduction of foods that may becontaminatedwithfecalbacteriaanddirectcontactwithhumanoranimal faeceswhenthe infant starts to grow. This age bracketalso represent a very active stage in thegrowth and development of children, thuscorresponding to the period when thechildren’s contact with the environmentalpathogensincreasesdramatically36,typicallydue to ignorance on the rudiments ofaseptic or hygienic practices.40 All these,including the high virulence of Salmonellaand its low infective dose make exposedchildren more vulnerable to infections.Detection of Salmonella diminished in theagebracket37-48months(19.2%).Thismaysignifybothanimprovementintheimmunestatus of the children and their eatinghabits.Mostentericpathogensstimulateatleast partial immunity against repeatedinfection or illness, which helps to explainthe declining incidence of disease in olderchildren and adults. The prevalence ofdiarrhea is also shown to generally reduceasthechildrengrowduetothesefactors.38

Low prevalence in the 0-12months of agegroup could be due to breast-feeding.Antibodiesinbreastmilkprotectthemfrominfectious agents. Bacteria like theSalmonella species, are associated withdairy products, fecally contaminated foodor water, hence proper hygiene andsanitation must be practiced to reduceinfectionbythesepathogens.Similartrendsinagerelatedprevalencewereobserved inotherreports.24,25,39Male children were more infected thantheir female counterparts. Female childrenwithin the study area are more protectedthanthemales;thiscouldalsoincreasethechances of infection in the male children.Abdullahi et al observed an overallSalmonella prevalence of 13.67% from a

study in Kano, Nigeria, out of which 8.0%weremaleand5.67%werefemale.39Similarobservations were made in studies byAdkins and Santiago,41 Al-Jurayyan,42 andNgoziandOnyekwe.43

The use of antimicrobial agents in thetreatment of diarrhea cannot beoveremphasized, because specificantimicrobialtreatmentmayberequiredtosupplement supportive anti-dehydrationtreatment, which is the cornerstone oftherapy of acute infantile diarrhea.However, non-adherence to treatmentstrategiesanddubiousdrugqualityandself-medication all favor the emergence ofmicrobial resistance.44 This was clearlyevident from this study where over 90%resistance to three antibiotics (Ampicillin,Amoxicilin, and Amoxicilin Clavulanic acid)bySalmonellaspecieswasobserved.It has become increasingly important tomonitor patterns of resistance as theantibacterial susceptibility of bacterialpathogenswhich contribute significantly totheburdenofinfantilediarrheaisdeclining.TheresistanceoftheSalmonella isolatestothe antibiotics tested in this study calls forgreat concern, as it depicts a highprevalence of antibiotic resistance bySalmonella strains to the β-lactam class ofantibiotics (Ampicillin, Amoxycillin,Amoxycillin-clavulanicacid,andCefotaxime)which are frequently used empirically forthetreatmentofdiarrheaandanumberofinfectious diseases. Similar trends inantibiotic resistance have been reportedpreviously.22,34,45,46

According to Ajibade,47 the susceptibilitypatterns reported in 2009 from differentlocationsinEkitiStateshowedresistancetostreptomycin, chloramphenicol, cefepime,nalidixicacidtetracyclineandtrimethoprim-sulfamethoxazole. This same trend ofresistancewas repeated in2010and2011,however, the notable change was asignificant increase in the resistance toampicillin from 20% to 100% in Ado Ekiti



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andfrom39%to100%atIkoleEkitiandalsoGentamicinfrom18%atIkere-Ekitiin2009,to 86% and 84% in 2010 and 2011respectively.However, all the strains had varyingpercentage susceptibility to Nalidixic acid(45.5%), Ciprofloxacin (90.9%), Cefotaxime(90.9%) and Chloramphenicol (63.3%). Thismeansthatthesechemotherapeuticagentsare effective in the treatment of diarrheacaused by these pathogens. The use ofCiprofloxacin in young children however,has grave risks as complications involvingtroubled breathing, swelling of the face,lips, tongue,mouth, or throat, irregular oruneven heartbeat, fainting or seizures candevelopassideeffects48.Resistance, particularly to the commonlyavailable antibiotics poses major healthconcerns because the most effectivechemotherapeutic agents such asCiprofloxacin are not readily available inmostruralandurbancommunities.Salmonella strains in this study generallyshowed 100% susceptibility to theaminoglycosidegentamicin,however, ithasbeenreported that invitro susceptibilityofSalmonella species and Shigella species toaminoglycosides may appear active.However, are not effective clinically andshouldnotbereportedassusceptible.21Theinfluence of antimicrobial resistance inSalmonella species is quite extensive,reaching many areas. Acquisition ofresistance genes adds complexity tolaboratory diagnosis and complicatestherapeutic outcomes. Antimicrobialresistance also affects the therapeuticregimen, leading to considerable publichealth concerns and substantial economicburden.Limitations: Some important aspects suchas socio-economic, and environmentalfactors coupled with health, physiologicaland behavioral risk factors could not becoveredinthestudy.

CONCLUSIONThese research findings show that, thoughthere are a number of causative agents ofdiarrheal diseases, Salmonella still remainsone of the major and most importantbacterial pathogen of diarrhea amongchildren in the study area. Age of childrenplays a significant role in both theirexposure and susceptibility of the studysubjects to Salmonella infections. Theresults of antibiotic susceptibility in thisstudy shows a high resistance rate amongisolatesespecially to theβ-lactamgroupofantibiotics making them completelyunreliableinthemanagementofSalmonellainfections in the study area. Multipleantibiotic resistancewas observed in 100%of the isolates, this defines the level ofsignificance antibiotic resistance hasbecome to public health especially in childhealth.Recommendations: As a public healthmeasure to reduce the disease burden, anintegrated package of immunizationservicesandotherchildcareprogramsneedto be implemented in addition to wellfocused health education messages toimprove treatment–seeking behavior forchildhood diarrhea as well as improvedpersonal and environmental hygiene. Theneed to continue to carry out extensivemulti-center studies involving both ruraland urban areas to identify all the riskfactors precipitating diarrhea will lead topoliciesonpreventiveprogramsglobally.Authoraffiliations1Department of Biological Sciences, FederalUniversity,Dutsin-Ma.KatsinaState,Nigeria2Department of Microbiology, Ahmadu BelloUniversity,Zaria.KadunaState,NigeriaREFERENCES1. Sánchez J, Holmgren J. Virulence

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