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Isolation and molecular characterization of Mycoplasma spp. in sheep and goats in Egypt.

ABSTRACT: Background and Aim:Different species of Mycoplasma are associated with many pathological problems in small ruminants including respiratory manifestation, this problem results in significant losses, especially in African countries. This study aimed to (I) study some epidemiological aspects of Mycoplasma species infections in Egyptian sheep and goats at Giza Governorate, (II) diagnosis of Mycoplasma species affections using bacterial isolation and identification, (III) apply the polymerase chain reaction (PCR) for typing of different Mycoplasma species, and (IV) illustrate the phylogenetic tree for the isolated Mycoplasma species and other species from GenBank using the purified PCR product. Materials and Methods:A total of 335 samples were collected from sheep and goats from Giza Governorate in Egypt as 142 nasal swabs from clinically affected animals, 167 pneumonic lungs, 18 samples from tracheal bifurcation, and 8 samples by bronchial wash were cultured on pleuropneumonia-like organisms (PPLOs) media for cultivation of Mycoplasma species. PCR and sequencing and phylogenetic analysis were adopted to identify and classify the isolated Mycoplasma species. Results:A total of 24 Mycoplasma isolates were isolated on PPLO media, identified by biochemical tests, and confirmed and typed by PCR using specific primers. 10 isolates were confirmed as Mycoplasma arginini, four isolates as Mycoplasma ovipneumoniae by PCR, and 10 isolates as undifferentiated Mycoplasma species. A purified isolate of M. arginini and M. ovipneumoniae was sequenced and phylogenetic analysis was illustrated. Conclusion:M. arginini and M. ovipneumoniae are prevalent in Egyptian sheep and goats. Further studies on M. arginini are required due to its high frequency of isolation from pneumonic sheep and goats and also from animals suffer from different respiratory manifestations.

SUBMITTER: Halium MMA

PROVIDER: S-EPMC6584854 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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Isolation and molecular characterization of <i>Mycoplasma</i> spp. in sheep and goats in Egypt.

Halium Mounier M Abdel MMA Salib Fayez A FA Marouf S A SA Massieh Emil S Abdel ESA

Veterinary world 20190513 5

<h4>Background and aim</h4>Different species of <i>Mycoplasma</i> are associated with many pathological problems in small ruminants including respiratory manifestation, this problem results in significant losses, especially in African countries. This study aimed to (I) study some epidemiological aspects of <i>Mycoplasma</i> species infections in Egyptian sheep and goats at Giza Governorate, (II) diagnosis of <i>Mycoplasma</i> species affections using bacterial isolation and identification, (III) ...[more]

PMID: 31327901

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Project description:Hemotropic mycoplasmas (hemoplasmas) are emerging zoonotic pathogens with a worldwide distribution that can cause mild to severe hemolytic anemia, icterus, ill-thrift, infertility, and poor weight gain. However, understanding of the molecular epidemiology of hemoplasmas (Mycoplasma ovis and 'Candidatus Mycoplasma haemovis') is limited in sheep and goats, and the hemoplasma strain/species/variant 'Candidatus M. haemovis' was poorly studied throughout the world and had never been detected in China until now. Thus, the aim of the present study was to determine the molecular prevalence of hemoplasmas, including M. ovis and 'Candidatus M. haemovis' in sheep and goats from seven provinces and one autonomous region of China.A total of 1364 blood samples were collected from sheep and goats in seven provinces and one autonomous region of China. All blood samples were tested for hemoplasmas (M. ovis and 'Candidatus M. haemovis') by nested PCR amplification based on 16S rRNA gene. Positive specimens underwent nucleotide sequencing and phylogenetic analysis.Overall, 610 specimens (44.7%, 610/1364) were shown to be hemoplasmas (M. ovis and 'Candidatus M. haemovis') -positive by nested PCR amplification based on 16S rRNA gene. The prevalence in goats was 44.1% (379/860), and 45.8% (231/504) in sheep, while that in grazing small ruminants was 54.4% (396/728) and 33.6% (214/636) in house feeding small ruminants. Sequencing of the nearly complete 16S rRNA gene was successful for the 103 randomly selected positive specimens from different farms in different sampling sites of China. Among them, analysis of the 16S rRNA gene sequences identified M. ovis (n = 56) and 'Candidatus M. haemovis' (n = 47). Two (KU983740 and KU983746) of the four novel genotypes obtained in this study were closely related to M. ovis, while the other two genotypes (KU983748 and KU983749) had high identity with 'Candidatus M. haemovis'. Remarkably, the genotype (KU983740) of M. ovis in sheep and goats in this study fell in a clade with two human hemoplasmas from USA (KF313922 and GU230144) and shared 99.8%-99.9% with them.In this study, 'Candidatus M. haemovis' was first detected in Chinese sheep and goats and hemoplasmas (M. ovis and 'Candidatus M. haemovis') are highly prevalent, and widely distributed in China.

Project description:BackgroundOrf is a highly contagious viral skin disease in sheep and goats caused by Orf virus (ORFV) in the genus Parapoxvirus. Although sheep and goats are considered an essential food resource, particularly in Africa, ORFV infection represents an increasing challenge to animal productivity causing high economic losses.AimThis study aimed to detect and characterize the ORFV in suspected clinically diseased goats in two neighboring Egyptian governorates, Al-Sharkia and Ismailia, flocks during April 2020 and July 2021by using PCR and phylogenetic analysis of partial B2L sequence.he present study indicate the necessity for establishing normal heart values in conscious and anaesthetized individuals.MethodsKids from two Egyptian governorates showed the clinical picture of ORFV infection. Samples were collected (n = 15) from two different flocks during April 2020 and July 2021. PCR was carried out to detect the ORFV by targeting a highly conserved sequence within ORFV (B2L) gene. To determine the phylogenetic relationship with other ORFV strains, sequencing and phylogenetic analysis were performed.ResultsORFV infection was confirmed in 12 samples of oral scabs (80%) by PCR targeting a highly conserved sequence within B2L gene. Sequencing of DNA products was performed and obtained sequences revealed 100% identity at the nucleotide level. Two ORFVs, one from each outbreak showed 98.2% nucleotide identity with a previous Egyptian ORFV (KP984529) whereas our isolates showed higher nucleotide identities, 99.1% and 98.7% with ORFV strains from neighboring countries, Sudan and Ethiopia, respectively. The phylogenetic tree grouped isolates into two main clusters, cluster I included isolates of this study and foreign ones mainly from China, India, and Sudan. Interestingly, the vaccine strains of ORF used in different countries were grouped in cluster II with previous Egyptian isolate (KP984529), Ethiopian and Israeli ORFV isolates.ConclusionMolecular characterization of B2L gene of ORFV isolates revealed higher sequence identities and more close genetic relationships with other ORFV strains circulating in neighboring countries than with the Egyptian isolates. These findings provide an insight into the genetic diversity of field ORFV isolates circulating in goats in the Egyptian governorates.

Project description:BackgroundIn Egypt, liver flukes, Fasciola spp. (Digenea: Fasciolidae), have a serious impact on the farming industry and public health. Both Fasciola hepatica and Fasciola gigantica are known to occur in cattle, providing the opportunity for genetic recombination. Little is known on the identity and genetic variability of Fasciola populations in sheep.MethodsThis study was performed to determine the prevalence of liver flukes in sheep in Menofia Province as a representative area of the delta region in Egypt, as measured by postmortem examination of slaughtered animals at three abattoirs. The identity and genetic variability of Fasciola spp. in slaughtered animals were determined by PCR-sequence analysis of the nuclear ribosomal internal transcribed spacer 1 (ITS1) and the mitochondrial NADH dehydrogenase subunit 1 (nad1) genes.ResultsPhysical inspection of the liver indicated that 302 of 2058 (14.7%) slaughtered sheep were infected with Fasciola spp. Sequence analysis of the ITS1 and nad1 genes of liver flukes from 17 animals revealed that 11 animals were infected with F. hepatica, four with F. gigantica, and two with both species. Seventy eight of 103 flukes genetically characterized from these animals were F. hepatica, 23 were F. gigantica, and two had ITS1 sequences identical to F. hepatica but nad1 sequences identical to F. gigantica. nad1 sequences of Egyptian isolates of F. gigantica showed pronounced differences from those in the GenBank database. Egyptian F. gigantica haplotypes formed haplogroup D, which clustered in a sister clade with haplogroups A, B and C circulating in Asia, indicating the existence of geographic isolation in the species.ConclusionsBoth F. hepatica and F. gigantica are prevalent in sheep in Egypt and an introgressed form of the two occurs as the result of genetic recombination. In addition, a geographically isolated F. gigantica population is present in the country. The importance of these observations in epidemiology of fascioliasis needs to be examined in future studies.

Dataset Information

Isolation and molecular characterization of Mycoplasma spp. in sheep and goats in Egypt.

Publications

Isolation and molecular characterization of <i>Mycoplasma</i> spp. in sheep and goats in Egypt.

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