208759

8102019 208759

httpslidepdfcomreaderfull208759 16

Research ArticleThe Zoonotic Risk of Ancylostoma ceylanicum Isolated fromStray Dogs and Cats in Guangzhou South China

Yuanjia Liu1 Guochao Zheng1 Muhamd Alsarakibi1 Xinheng Zhang2

Wei Hu1 Liqin Lin1 Liping Tan1 Qin Luo1 Pengyun Lu1 and Guoqing Li1

983089 College of Veterinary Medicine South China Agricultural University Guangzhou 983093983089983088983094983092983090 China983090 College of Animal Science South China Agricultural University Guangzhou 983093983089983088983094983092983090 China

Correspondence should be addressed to Guoqing Li gqliscaueducn

Received 983096 February 983090983088983089983092 Accepted 983090983090 April 983090983088983089983092 Published 983092 May 983090983088983089983092

Academic Editor Francesca Mancianti

Copyright copy 983090983088983089983092 Yuanjia Liu et al Tis is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Canine and eline hookworm inection is endemic in many countries with zoonotic transmission representing a potentially signi1047297cant public health concern However there is limited data available on the zoonotic transmission o canine and elinehookworms in China Tis study was conducted to evaluate the zoonotic risk o Ancylostoma ceylanicum isolated rom stray dogs and cats in Guangzhou south China Primer pairs CAFCAR were designed to ampliy complete IS sequences o obtained A ceylanicum Te results were compared with ourteen IS reerence sequences o human-derived A ceylanicum registered in

GenBank and phylogenetic trees were established by using NJ and ML methods Te sequence similarity o three dog-derivedand 1047297ve cat-derived A ceylanicum with ourteen human-derived A ceylanicum were 983097983094983096sim983089983088983088 and 983097983095983096sim983089983088983088 respectivelyPhylogenetic analysis placed A ceylanicum isolated rom dogs and cats in the same group with A ceylanicum human isolatesDue to the ability o A ceylanicum to cause a patent inection in humans the zoonotic risk arising rom dog and cat reservoirs tocommunities in this region should be determined

1 Introduction

Hookworms are one o the most common parasitic nema-todes which inect mammals including humans dogs cats

and wild canids [983089] Tey are prevalent in temperate and trop-ical regions o the world [983090] Currently more than 983089983088983088 speciesbelonging to 983089983096 genera o hookworms have been discoveredparasitizing the intestinal tract o carnivore and herbivoreanimal hosts

Ancylostoma ceylanicum is the only species o canineand eline hookworms which can develop into adult orm inthe human intestinal tract causing iron-de1047297ciency anemia[983091] Parasitic to cats and dogs A ceylanicum is inective tohumans as a zoonosis in some regions o Asia but has notbeen associated with host blood loss in humans [983092] andthereore was not considered a major pathogen [983093] In Ma-laysia the prevalence o A ceylanicum in humans was 983090983091983092

with A ceylanicum being the secondmost common species o hookworms [983091] while one-third o human hookworm inec-tion was caused by A ceylanicum in Laos [983094] A ceylanicumwas isolated rom both humans and dogs in Tailand as well

and only in cases o human inection dyspepsia abdominalpain diarrhea and other gastrointestinal symptoms wasreported [983095] Tere were also reports o human inection with

A ceylanicum in aiwan [983096] and Fujian [983097] In Guangzhouthe inection rate o A ceylanicum in stray dogs and catswas 983092983090983094983095 [983089983088] and 983092983088983090 [983089983089] respectively Consideringthe zoonotic potential o A ceylanicum inection betweenanimals and humans it is very important to evaluate thezoonotic risk o A ceylanicum Te aims o this study were toampliy the complete IS sequences o A ceylanicum isolatedrom dogs and cats in Guangzhou to compare the sequenceso dog-derived and cat-derived A ceylanicum with human-derived A ceylanicum and to construct a phylogenetic tree

Hindawi Publishing CorporationBioMed Research InternationalVolume 2014 Article ID 208759 5 pageshttpdxdoiorg1011552014208759

8102019 208759


983090 BioMed Research International

using ML and NJ methods to evaluate the zoonotic risk o Aceylanicum between pet (dogs and cats) and humans

2 Materials and Methods

983090983089 Egg Samples Egg samples o A ceylanicumwerecollected

through an epidemiologic survey o hookworm inections indogs ( = 254 Guangzhou) and cats ( = 102 Guangzhou)in two previous studies [983089983088 983089983089] preserved in 983090983093 potassiumdichromate and stored at 983092∘C

983090983090 Genomic DNA Extraction Glass beads and proteinase Kwere added to 983089983093 mL tube containing egg samples o A cey-lanicum Te suspension was thoroughly mixed with a vortexmixer or 983091983088 minutes and then incubated at 983093983093∘C or 983089983094 hours[983089983089] DNA puri1047297cation was carried out according to the man-uacturerrsquos protocols

983090983091 PCR Ampli1047297cation of the Complete IS Sequence Tecomplete IS was ampli1047297ed using a orward primer CAF(983093-GACGCGGAC GCGA-983091) and a reverse primerCAR (983093-AAGCAGCGGGAGCA-983091) [983089983089] that werederived rom A ceylanicum (GenBank JQ983096983089983090983094983097983092 AJ983097983090983088983091983092983095AM983088983091983097983095983091983097) and A ceylanicum (GenBank DQ983091983096983089983093983092983089DQ983095983096983088983088983088983097) Te PCR ampli1047297cation was perormed in 983090983093 1038389L

volume containing 983089983095983091 1038389L o distilled water 983090983093 1038389L o 983089983088times PCR Buffer 983090 1038389L o diethylnitrophenyl thiophosphate(dNP 983090983093 mM each) 983088983093 1038389L o each primer (CAFCAR983093983088 pmoluL) 983088983090 1038389L o Ex-aq polymerase (983093 U1038389L) and983090 1038389Lo the DNA sample Te PCR reactions applied ollowingcycling pro1047297le initial denaturation o 983097983094∘C or 983093 minutes ol-lowed by 983091983093 cycles o denaturation annealing and extensionat 983097983094∘C or 983091983088 seconds 983094983088∘C or 983091983088 seconds and 983095983090∘C or 983094983088seconds respectively and the 1047297nal extension was carried outat 983095983090∘C or 983095 minutes Te PCR products were analyzed with983089983088 agarose gel electrophoresis at voltage 983089983090983088 V or about983090983088 minutes Te results on the gel were observed under theUV transilluminator and recorded

983090983092 Puri1047297cation and Sequencing of PCR Products Te posi-tive PCR products by agarose gel electrophoresis were puri-1047297ed using UNIQ-983089983088 column Cyclo-Prep PCR Clean-Up KitTe puri1047297ed products were sequenced by Huada Gene Bio-technology Co Ltd (Guangzhou)

983090983093 Similarity Analysis of IS Sequences Homology analysiso A ceylanicum IS sequences rom dogs andcats with our-teen IS sequences o human-derived A ceylanicum (Gen-Bank JF983097983094983088983091983094983090 JF983097983094983088983091983094983091 JF983097983094983088983091983094983093 JF983097983094983088983091983094983095 JF983097983094983088983091983094983096JF983097983094983088983091983094983097 JN983089983090983088983096983095983089 JN983089983090983088983096983095983090 JN983089983090983088983096983095983092 JN983089983090983088983096983095983093JN983089983090983088983096983095983094 JN983089983090983088983096983095983095 JN983089983090983088983096983096983088 and JN983089983090983088983096983096983089) was done us-ing the MegAlign program o DNAStar (version 983095 MadisonWI USA)

983090983094 Phylogenetic Analysis Based on IS Sequences Te ISsequences were aligned using the computer program CLUS-AL X 983089983096983089 Phylogenetic trees were constructed by using

1 2 3 4 5 6 7 8

5000

3000

2000

1000

750

F983145983143983157983154983141 983089 PCR ampli1047297cation o IS sequences o Ancylostomaceylanicum romdogs and cats Numbers 983089ndash983096 represent eightisolatesG983090983091 G983091983090 G983090983089 M983094 M983093983093 M983093983096 M983094983088 and M983095983094 respectively

neighbor-joining (NJ) and maximum likelihood (ML) meth-ods employing MEGA 983093983089 sofware (Arizona State UniversityUSA) wo methods were carried out under deault settingand the consensus tree was obtained afer bootstrap analysiswith 983089983088983088983088 replications

3 Results and Discussion

983091983089 PCR Ampli1047297cation and Sequencing of Complete IS Se-quence PCR ampli1047297cation o the IS yielded a ragment o 983097983088983088 bp (Figure 983089) and subsequent sequencing con1047297rmed thatthe ragment contained partial 983089983096S rDNA sequence completeIS sequence and partial 983090983096S rDNA sequence Te completeIS sequence o three dog-derived A ceylanicum isolates(G983090983091 G983091983090 and G983090983089) and 1047297ve cat-derived A ceylanicumisolates (M983094 M983093983093 M983093983096 M983094983088 and M983095983094) was 983095983091983096 bp inlength containing IS983089 (983091983094983092 bp) 983093983096S rDNA (983089983093983091 bp) andIS983090 (983090983090983089 bp) Te eight sequences had been submitted to

GenBank (KC983095983093983093983088983090983095 KF983090983095983097983089983091983090mdashKF983090983095983097983089983091983096) respectively

983091983090 Similarity of IS Sequences BLAS results o dog-derived A ceylanicum isolates (G983090983091 G983091983090 and G983090983089) showed983097983094983096sim983089983088983088 homology with 983089983092 human-derived A ceylan-icum where the homology o two dog isolates (G983090983091 G983091983090)with one human-derived A ceylanicum isolate (GenBankJN983089983090983088983096983095983093) was 983097983094983096 while one dog isolate (G983090983089) was 983089983088983088homologous to three human-derived A ceylanicum (Gen-Bank JF983097983094983088983091983094983090 JF983097983094983088983091983094983096 and JN983089983090983088983096983095983089) (Figure 983090)

BLAS results rom IS sequences o 1047297ve cat-derived Aceylanicum isolates (M983094 M983093983093 M983093983096 M983094983088 and M983095983094) were983097983095983096sim983089983088983088 homologous with 983089983092 human-derived A ceylan-

icum isolates Only the isolate (M983093983093) was 983097983095 homologouswith the human-derived A ceylanicum isolate (GenBankJN983089983090983088983096983095983093) while several cat-derived A ceylanicum isolateswere 983089983088983088 homologous with human-derived A ceylanicumisolates (eg M983093983093 with JN983089983090983088983096983095983095 M983093983096 with JN983089983090983088983096983095983089JF983097983094983088983091983094983096 JF983097983094983088983091983094983090 M983094 with JF983097983094983088983091983094983096 JN983089983090983088983096983095983089 JF983097983094983088983091983094983090M983094983088 with JN983089983090983088983096983095983095 and M983095983094 with JF983097983094983088983091983094983090 JF983097983094983088983091983094983096 andJN983089983090983088983096983095983089) (Figure 983090)

983091983091 Phylogenetic Analysisof IS Sequence Phylogenetic anal-ysis o obtained IS sequences whether by NJ method or MLmethod revealed a generally consistent topological structureo a phylogenetic tree it showed that the clades o the six

8102019 208759


8102019 208759



Human A ceylanicum JF960369

Dog A ceylanicum G32

Human A ceylanicum JN120881









Cat A ceylanicum M58 A ceylanicum


Cat A ceylanicum M60






Cat A ceylanicum M6



A brazilienseU stenocephala

A duodenale

A tubaeforme

A caninum

0

6464

58

73

6746

37

0002

(a)

A ceylanicum






Cat A ceylanicum M6














A duodenale

U stenocephala

A braziliense

0 01




A caninum

A tubaeforme

(b)

F983145983143983157983154983141 983091 Phylogenetic relationship o hookworms rom pets (dogs and cats) and humans based on IS sequence by NJ (a) and ML (b)methods

risk arising rom dog andcat reservoirs to communities in thetropical regions should be determined Future studies should

also be carried out on animals to determine the extent o Aceylanicum inection in Guangzhou and to assess the risk o zoonotic transmission and disease

Conflict of Interests

Te authors declare that there is no con1047298ict o interestsregarding the publication o this paper

Acknowledgments

Tis work was unded by the National Natural Science

Foundation o China (Grant nos 983091983088983097983095983090983089983095983097 983091983089983090983095983090983093983093983089) Teauthors would like to thank Xu-hui Lin and the humaneshelterrsquos personnel or helping them collect all o the samples

References

[983089] F A Smout R C Tompson and L F Skerrat ldquoFirst report o Ancylostoma ceylanicum in wild canidsrdquo International Journal for Parasitology Parasites and Wildlife vol 983090 pp 983089983095983091ndash983089983095983095 983090983088983089983091

[983090] N R de Silva S Brooker P J Hotez A Montresor D Engelsand L Savioli ldquoSoil-transmitted helminth inections updatingthe global picturerdquo rends in Parasitology vol983089983097no983089983090pp983093983092983095ndash983093983093983089 983090983088983088983091

[983091] R Ngui Y A L Lim R raub R Mahmud and M S MistamldquoEpidemiological and genetic data supporting the transmission

o Ancylostoma ceylanicum among human and domestic ani-malsrdquo PLoS Neglected ropical Diseases vol 983094 no 983090 Article IDe983089983093983090983090 983090983088983089983090

[983092] S M Carroll and D I Grove ldquoExperimental inection o humans with Ancylostoma ceylanicum clinical parasitologicalhaematological and immunological 1047297ndingsrdquo ropical and Geo- graphical Medicine vol 983091983096 no 983089 pp 983091983096ndash983092983093 983089983097983096983094

[983093] P J Hotez J M Hawdon M Cappello B F Jones and DI Pritchard ldquoMolecular pathobiology o hookworm inectionrdquoInfectious Agents and Disease vol 983092 no 983090 pp 983095983089ndash983095983093 983089983097983097983093

[983094] J V Conlan B Sripa S Attwood and P N Newton ldquoA review o parasitic zoonoses in a changing Southeast Asiardquo Veterinary Parasitology vol 983089983096983090 no 983089 pp 983090983090ndash983092983088 983090983088983089983089

[983095] R J raub Inpankaew C Sutthikornchai Y Sukthana

and R C A Tompson ldquoPCR-based coprodiagnostic toolsreveal dogs as reservoirs o zoonotic ancylostomiasis caused by Ancylostoma ceylanicum in temple communities in BangkokrdquoVeterinary Parasitology vol 983089983093983093 no 983089-983090 pp 983094983095ndash983095983091 983090983088983088983096

[983096] Y C Hsu and J Lin ldquoImages in clinical medicine Intestinalinestation with Ancylostoma ceylanicumrdquo Te New England Journal of Medicine vol 983091983094983094 no 983089983091 article e983090983088 983090983088983089983090

[983097] Q Q Chen X M Lin and C C Yang ldquoA biology andepidemiological study o Ancylostoma ceylanicum 1047297rst oundin human and Felis bengalensis in Fujian Province o Chinardquo Journal of Xiamen University vol 983090983088 no 983092 pp 983092983095983094ndash983092983096983094 983089983097983096983089

[983089983088] Y J Liu G C Zheng P Zhang et al ldquoMolecular identi1047297cationo hookworms in stray and shelter dogs rom Guangzhou CityChina using IS sequencesrdquo Journal of Helminthology 983090983088983089983091

8102019 208759


BioMed Research International 983093

[983089983089] Y Liu G C Zheng M Alsarakibi et al ldquoMolecular identi1047297-cation o Ancylostoma caninum isolated rom cats in southernChina based on complete IS sequencerdquo BioMed ResearchInternational vol 983090983088983089983091 Article ID 983096983094983096983088983093983088 983094 pages 983090983088983089983091

[983089983090] D D Bowman S P Montgomery A M Zajac M L Eberhardand K R Kazacos ldquoHookworms o dogs and cats as agents o cutaneous larva migransrdquo rends in Parasitology vol 983090983094 no 983092pp 983089983094983090ndash983089983094983095 983090983088983089983088

[983089983091] R J raub R P Hobbs P J Adams J M Behnke P D Har-ris and R C A Tompson ldquoA case o mistaken identitymdashreappraisal o the species o canid and elid hookworms (Ancy-lostoma) present in Australia and Indiardquo Parasitology vol 983089983091983092no 983089 pp 983089983089983091ndash983089983089983097 983090983088983088983095

[983089983092] P J Hotez and A Kamath ldquoNeglected tropical diseases in sub-Saharan Arica review o their prevalence distribution anddisease burdenrdquo PLoS Neglected ropical Diseases vol 983091 no 983096article e983092983089983090 983090983088983088983097

[983089983093] S Y Zhuang and D X Jin ldquoInvestigation on hookworm inec-tion in carnivorous animals in Guizhou Province and researchon the lie history o Ancylostoma ceylanicumrdquo Journal of

Guiyang Medical College vol 983095 pp 983093983092ndash983094983097 983089983097983096983090 (Chinese)[983089983094] C S Palmer R J raub I D Robertson et al ldquoTe veterinary

and public health signi1047297cance o hookworm in dogs and cats inAustralia and the status o A ceylanicumrdquo Veterinary Parasitol-ogy vol 983089983092983093 no 983091-983092 pp 983091983088983092ndash983091983089983091 983090983088983088983095

8102019 208759


Submit your manuscripts at

httpwwwhindawicom

8102019 208759



using ML and NJ methods to evaluate the zoonotic risk o Aceylanicum between pet (dogs and cats) and humans

2 Materials and Methods

983090983089 Egg Samples Egg samples o A ceylanicumwerecollected

through an epidemiologic survey o hookworm inections indogs ( = 254 Guangzhou) and cats ( = 102 Guangzhou)in two previous studies [983089983088 983089983089] preserved in 983090983093 potassiumdichromate and stored at 983092∘C

983090983090 Genomic DNA Extraction Glass beads and proteinase Kwere added to 983089983093 mL tube containing egg samples o A cey-lanicum Te suspension was thoroughly mixed with a vortexmixer or 983091983088 minutes and then incubated at 983093983093∘C or 983089983094 hours[983089983089] DNA puri1047297cation was carried out according to the man-uacturerrsquos protocols

983090983091 PCR Ampli1047297cation of the Complete IS Sequence Tecomplete IS was ampli1047297ed using a orward primer CAF(983093-GACGCGGAC GCGA-983091) and a reverse primerCAR (983093-AAGCAGCGGGAGCA-983091) [983089983089] that werederived rom A ceylanicum (GenBank JQ983096983089983090983094983097983092 AJ983097983090983088983091983092983095AM983088983091983097983095983091983097) and A ceylanicum (GenBank DQ983091983096983089983093983092983089DQ983095983096983088983088983088983097) Te PCR ampli1047297cation was perormed in 983090983093 1038389L

volume containing 983089983095983091 1038389L o distilled water 983090983093 1038389L o 983089983088times PCR Buffer 983090 1038389L o diethylnitrophenyl thiophosphate(dNP 983090983093 mM each) 983088983093 1038389L o each primer (CAFCAR983093983088 pmoluL) 983088983090 1038389L o Ex-aq polymerase (983093 U1038389L) and983090 1038389Lo the DNA sample Te PCR reactions applied ollowingcycling pro1047297le initial denaturation o 983097983094∘C or 983093 minutes ol-lowed by 983091983093 cycles o denaturation annealing and extensionat 983097983094∘C or 983091983088 seconds 983094983088∘C or 983091983088 seconds and 983095983090∘C or 983094983088seconds respectively and the 1047297nal extension was carried outat 983095983090∘C or 983095 minutes Te PCR products were analyzed with983089983088 agarose gel electrophoresis at voltage 983089983090983088 V or about983090983088 minutes Te results on the gel were observed under theUV transilluminator and recorded

983090983092 Puri1047297cation and Sequencing of PCR Products Te posi-tive PCR products by agarose gel electrophoresis were puri-1047297ed using UNIQ-983089983088 column Cyclo-Prep PCR Clean-Up KitTe puri1047297ed products were sequenced by Huada Gene Bio-technology Co Ltd (Guangzhou)

983090983093 Similarity Analysis of IS Sequences Homology analysiso A ceylanicum IS sequences rom dogs andcats with our-teen IS sequences o human-derived A ceylanicum (Gen-Bank JF983097983094983088983091983094983090 JF983097983094983088983091983094983091 JF983097983094983088983091983094983093 JF983097983094983088983091983094983095 JF983097983094983088983091983094983096JF983097983094983088983091983094983097 JN983089983090983088983096983095983089 JN983089983090983088983096983095983090 JN983089983090983088983096983095983092 JN983089983090983088983096983095983093JN983089983090983088983096983095983094 JN983089983090983088983096983095983095 JN983089983090983088983096983096983088 and JN983089983090983088983096983096983089) was done us-ing the MegAlign program o DNAStar (version 983095 MadisonWI USA)

983090983094 Phylogenetic Analysis Based on IS Sequences Te ISsequences were aligned using the computer program CLUS-AL X 983089983096983089 Phylogenetic trees were constructed by using

1 2 3 4 5 6 7 8

5000

3000

2000

1000

750

F983145983143983157983154983141 983089 PCR ampli1047297cation o IS sequences o Ancylostomaceylanicum romdogs and cats Numbers 983089ndash983096 represent eightisolatesG983090983091 G983091983090 G983090983089 M983094 M983093983093 M983093983096 M983094983088 and M983095983094 respectively

neighbor-joining (NJ) and maximum likelihood (ML) meth-ods employing MEGA 983093983089 sofware (Arizona State UniversityUSA) wo methods were carried out under deault settingand the consensus tree was obtained afer bootstrap analysiswith 983089983088983088983088 replications

3 Results and Discussion

983091983089 PCR Ampli1047297cation and Sequencing of Complete IS Se-quence PCR ampli1047297cation o the IS yielded a ragment o 983097983088983088 bp (Figure 983089) and subsequent sequencing con1047297rmed thatthe ragment contained partial 983089983096S rDNA sequence completeIS sequence and partial 983090983096S rDNA sequence Te completeIS sequence o three dog-derived A ceylanicum isolates(G983090983091 G983091983090 and G983090983089) and 1047297ve cat-derived A ceylanicumisolates (M983094 M983093983093 M983093983096 M983094983088 and M983095983094) was 983095983091983096 bp inlength containing IS983089 (983091983094983092 bp) 983093983096S rDNA (983089983093983091 bp) andIS983090 (983090983090983089 bp) Te eight sequences had been submitted to

GenBank (KC983095983093983093983088983090983095 KF983090983095983097983089983091983090mdashKF983090983095983097983089983091983096) respectively

983091983090 Similarity of IS Sequences BLAS results o dog-derived A ceylanicum isolates (G983090983091 G983091983090 and G983090983089) showed983097983094983096sim983089983088983088 homology with 983089983092 human-derived A ceylan-icum where the homology o two dog isolates (G983090983091 G983091983090)with one human-derived A ceylanicum isolate (GenBankJN983089983090983088983096983095983093) was 983097983094983096 while one dog isolate (G983090983089) was 983089983088983088homologous to three human-derived A ceylanicum (Gen-Bank JF983097983094983088983091983094983090 JF983097983094983088983091983094983096 and JN983089983090983088983096983095983089) (Figure 983090)

BLAS results rom IS sequences o 1047297ve cat-derived Aceylanicum isolates (M983094 M983093983093 M983093983096 M983094983088 and M983095983094) were983097983095983096sim983089983088983088 homologous with 983089983092 human-derived A ceylan-

icum isolates Only the isolate (M983093983093) was 983097983095 homologouswith the human-derived A ceylanicum isolate (GenBankJN983089983090983088983096983095983093) while several cat-derived A ceylanicum isolateswere 983089983088983088 homologous with human-derived A ceylanicumisolates (eg M983093983093 with JN983089983090983088983096983095983095 M983093983096 with JN983089983090983088983096983095983089JF983097983094983088983091983094983096 JF983097983094983088983091983094983090 M983094 with JF983097983094983088983091983094983096 JN983089983090983088983096983095983089 JF983097983094983088983091983094983090M983094983088 with JN983089983090983088983096983095983095 and M983095983094 with JF983097983094983088983091983094983090 JF983097983094983088983091983094983096 andJN983089983090983088983096983095983089) (Figure 983090)

983091983091 Phylogenetic Analysisof IS Sequence Phylogenetic anal-ysis o obtained IS sequences whether by NJ method or MLmethod revealed a generally consistent topological structureo a phylogenetic tree it showed that the clades o the six

8102019 208759


8102019 208759






















Cat A ceylanicum M6




A duodenale

A tubaeforme

A caninum

0

6464

58

73

6746

37

0002

(a)

A ceylanicum






Cat A ceylanicum M6














A duodenale

U stenocephala

A braziliense

0 01




A caninum

A tubaeforme

(b)






Acknowledgments



References













8102019 208759










8102019 208759



httpwwwhindawicom

8102019 208759


8102019 208759






















Cat A ceylanicum M6




A duodenale

A tubaeforme

A caninum

0

6464

58

73

6746

37

0002

(a)

A ceylanicum






Cat A ceylanicum M6














A duodenale

U stenocephala

A braziliense

0 01




A caninum

A tubaeforme

(b)






Acknowledgments



References













8102019 208759










8102019 208759



httpwwwhindawicom

8102019 208759






















Cat A ceylanicum M6




A duodenale

A tubaeforme

A caninum

0

6464

58

73

6746

37

0002

(a)

A ceylanicum






Cat A ceylanicum M6














A duodenale

U stenocephala

A braziliense

0 01




A caninum

A tubaeforme

(b)






Acknowledgments



References













8102019 208759










8102019 208759



httpwwwhindawicom

8102019 208759










8102019 208759



httpwwwhindawicom

8102019 208759



httpwwwhindawicom

208759

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