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Molecular and phylogenetic study on Toxocara vitulorum from cattle in the mid-Delta of Egypt.

ABSTRACT: Toxocara vitulorum is an important intestinal nematode that commonly infects ruminants world-wide, notably in tropical and sub-tropical regions. In Egypt, T. vitulorum has a high prevalence rate in cattle and buffaloes calves. The current work aims to identify and verify T. vitulorum collected from cattle in El-Mahlla El-Kubra city in the mid-Delta of Egypt, using molecular and phylogenetic tools. The first internal transcribed spacer (ITS-1) and 18S genes of ribosomal DNA were amplified, sequenced, and compared with nucleotide sequences deposited in data bases, and also used to construct the phylogenetic trees. Our results confirm that T. vitulorum isolated from cattle in Egypt is genetically identical to those recorded in other countries. Moreover, the phylogenetic trees show a close relationship among different species of Toxocara, including the zoonotic species. Our results show that ITS genes can be targeted as genetic markers to diagnose and discriminate among different Toxocara spp. The data presented here may be helpful in the pursuit of further molecular and genetic studies of Toxocara species.

SUBMITTER: Sultan K

PROVIDER: S-EPMC4554590 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature

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Molecular and phylogenetic study on Toxocara vitulorum from cattle in the mid-Delta of Egypt.

Sultan Khaled K Omar Mosaab M Desouky Abdelrazek Y AY El-Seify Mahmoud A MA

Journal of parasitic diseases : official organ of the Indian Society for Parasitology 20131119 3

Toxocara vitulorum is an important intestinal nematode that commonly infects ruminants world-wide, notably in tropical and sub-tropical regions. In Egypt, T. vitulorum has a high prevalence rate in cattle and buffaloes calves. The current work aims to identify and verify T. vitulorum collected from cattle in El-Mahlla El-Kubra city in the mid-Delta of Egypt, using molecular and phylogenetic tools. The first internal transcribed spacer (ITS-1) and 18S genes of ribosomal DNA were amplified, sequen ...[more]

PMID: 26345077

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Project description:Toxocara vitulorum is an Ascarid nematode infecting the small intestine of buffalo and cattle particularly neonate calves, with the postnatal route through milk is the main infection source. However, little is known about shedding rates and the optimum detection methods of T. vitulorum larvae in the milk of the infected bovine hosts. In this study, we aimed to evaluate the use of two methods, microscopy and PCR, and their detection limits both under the experimental and natural infection situations. In doing this, T. vitulorum eggs extracted from naturally occurring adult female worms were successfully subjected to experimental embryonation, and larvae were implemented in experimental infection of milk in ascending infection doses of 0, 1, 5, 10, 20, 50 larvae/2-ml milk samples. With the except of negative control, microscopy-based examination detected larvae in all samples, albeit with means, ranges, and the total number of larvae were detected in exponential rates relative to larvae densities in milk samples. PCR technique corresponded well to microscopy in detecting genomic DNA of T. vitulorum larvae in all milk samples down to a single larva/sample. On the other hand, and by applying the same methodology approach on 50 naturally-occurring bovine colostrum/milk samples, 13 (26%) and 20 (40%) samples were tested positive for T. vitulorum infection by microscopy and the PCR-based detection, respectively. Of these, 11 out of 26 buffalo samples (42.30%) and 2 out of 24 cow samples (8.33%) were tested positive by microscopy, while 16 (61.54%) and 3 (12.50%) of buffalo and cow samples were tested positive by PCR, respectively. By applying the Agreement Coefficient, substantial agreement (0.77) between molecular and microscopy detection was detected from all tested samples. In conclusion, larvae of T. vitulorum were unequivocally detected by microscopy and molecular methods in milk samples both under the experimental and natural field situations. Nevertheless, slightly higher rates by PCR than microscopy were obtained when detecting naturally-infected milk samples. To the best of our knowledge, this is the first in situ detection of larvae of T. vitulorum in the milk of the naturally infected animals.

Project description:IntroductionIndustrial livestock farming is a possible source of multi-resistant Gram-negative bacteria, including producers of extended spectrum beta-lactamases (ESBLs) conferring resistance to 3rd generation cephalosporins. Limited information is currently available on the situation of ESBL producers in livestock farming outside of Western Europe. A surveillance study was conducted from January to May in 2014 in four dairy cattle farms in different areas of the Nile delta, Egypt.Materials and methodsIn total, 266 samples were collected from 4 dairy farms including rectal swabs from clinically healthy cattle (n = 210), and environmental samples from the stalls (n = 56). After 24 h pre-enrichment in buffered peptone water, all samples were screened for 3rd generation cephalosporin-resistant Escherichia coli using Brilliance™ ESBL agar. Suspected colonies of putatively ESBL-producing E. coli were sub-cultured and subsequently genotypically and phenotypically characterized. Susceptibility testing using the VITEK-2 system was performed. All suspect isolates were genotypically analyzed using two DNA-microarray based assays: CarbDetect AS-1 and E. coli PanType AS-2 kit (ALERE). These tests allow detection of a multitude of genes and their alleles associated with resistance toward carbapenems, cephalosporins, and other frequently used antibiotics. Serotypes were determined using the E. coli SeroGenotyping AS-1 kit (ALERE).ResultsOut of 266 samples tested, 114 (42.8%) ESBL-producing E. coli were geno- and phenotypically identified. 113 of 114 phenotypically 3rd generation cephalosporin-resistant isolates harbored at least one of the ESBL resistance genes covered by the applied assays [blaCTX-M15 (n = 105), blaCTX-M9 (n = 1), blaTEM (n = 90), blaSHV (n = 1)]. Alarmingly, the carbapenemase genes blaOXA-48 (n = 5) and blaOXA-181 (n = 1) were found in isolates that also were phenotypically resistant to imipenem and meropenem. Using the array-based serogenotyping method, 66 of the 118 isolates (55%) could be genotypically assigned to O-types.ConclusionThis study is considered to be a first report of the high prevalence of ESBL-producing E. coli in dairy farms in Egypt. ESBL-producing E. coli isolates with different underlying resistance mechanisms are common in investigated dairy cattle farms in Egypt. The global rise of ESBL- and carbapenemase-producing Gram-negative bacteria is a big concern, and demands intensified surveillance.

Dataset Information

Molecular and phylogenetic study on Toxocara vitulorum from cattle in the mid-Delta of Egypt.

Publications

Molecular and phylogenetic study on Toxocara vitulorum from cattle in the mid-Delta of Egypt.

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