Detection, characterization, and molecular typing of human Mycoplasma spp. from major hospitals in Cairo, Egypt.
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ABSTRACT:
SUBMITTER: Metwally MA
PROVIDER: S-EPMC4258911 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Background and aimDifferent 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 methodsA 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.ResultsA 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.ConclusionM. 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.
| S-EPMC6584854 | biostudies-literature
Project description:Mycoplasma spp. are wall-less bacteria able to infect mammals and are classified as hemotropic (hemoplasma) and nonhemotropic. In aquatic mammals, hemoplasma have been reported in California sea lions (Zalophus californianus) and river dolphins (Inia spp.). We investigated Mycoplasma spp. in blood samples of West Indian manatees (Trichechus manatus), pinnipeds (5 species), and marine cetaceans (18 species) that stranded or were undergoing rehabilitation in Brazil during 2002-2022. We detected Mycoplasma in blood of 18/130 (14.8%) cetaceans and 3/18 (16.6%) pinnipeds. All tested manatees were PCR-negative for Mycoplasma. Our findings indicate that >2 different hemoplasma species are circulating in cetaceans. The sequences from pinnipeds were similar to previously described sequences. We also detected a nonhemotropic Mycoplasma in 2 Franciscana dolphins (Pontoporia blainvillei) that might be associated with microscopic lesions. Because certain hemoplasmas can cause disease and death in immunosuppressed mammals, the bacteria could have conservation implications for already endangered aquatic mammals.
| S-EPMC10683811 | biostudies-literature
Project description:BACKGROUND:The transmission of Cryptosporidium spp. and Giardia duodenalis into humans varies according to species/genotypes of the pathogens. Although infections with both parasites are recorded in Egypt, few data are available on the distribution of Cryptosporidium species and G. duodenalis genotypes. The present study assessed the occurrence and genetic diversity of Cryptosporidium spp. and G. duodenalis in Egyptian children. METHODS:In the present study, 585 fecal specimens were collected from children eight years old and younger in three provinces (El-Dakahlia, El-Gharbia and Damietta) during March 2015 to April 2016. PCR-RFLP analysis of the small subunit rRNA gene and sequence analysis of the 60 kDa glycoprotein gene were used to detect and subtype Cryptosporidium spp., respectively, whereas PCR and sequence analyses of the triose phosphate isomerase, glutamate dehydrogenase and ?-giardin genes were used to detect and genotype Giardia duodenalis. RESULTS:The overall infection rates of Cryptosporidium spp. and G. duodenalis were 1.4% and 11.3%, respectively. The Cryptosporidium species identified included C. hominis and C. parvum, each with three subtype families. The C. hominis subtypes were IbA6G3 (n = 2), IdA17 (n = 1), IdA24 (n = 1) and IfA14G1R5 (n = 1), while C. parvum subtypes were IIdA20G1 (n = 1), IIaA15G2R1 (n = 1), and IIcA5G3a (n = 1). The G. duodenalis identified included both assemblages A (n = 31) and B (n = 34). All G. duodenalis assemblage A belonged to the anthroponotic sub-assemblage AII, while a high genetic heterogeneity was seen within assemblage B. CONCLUSIONS:Data from this study are useful in our understanding of the genetic diversity of Cryptosporidium spp. and G. duodenalis in Egypt and the potential importance of anthroponotic transmission in the epidemiology of both pathogens.
| S-EPMC6042380 | biostudies-literature
Project description:Cattle are intermediate hosts for several Sarcocystis species with different definitive hosts. The present study, to our knowledge, is the first to determine the prevalence of Sarcocystis infection and morphologically and molecularly identify Sarcocystis species in cattle in Qena Governorate, Upper Egypt. The samples were collected from the heart and oesophagus muscles of 84 slaughtered cattle (76 males and 8 females) aged between 11 months and 3 years from slaughterhouses in different localities in Qena Governorate, Upper Egypt. The samples were macroscopically and histologically examined, and the molecular identification of the species was performed using 18S ribosomal subunit DNA through PCR and DNA sequencing. Infection was detected in 72 out of 84 animals (85.7%) and was more prevalent in males (76.2%) than in females (9.5). Using light microscopy, the microscopic sarcocysts were observed to be thin-walled. Sequencing and genotyping revealed one isolate that had 99 and 100% identity, respectively, to Sarcocystis cruzi, while another isolate had 95 and 99% identity to Sarcocystis hjorti. The present study is the first to identify Sarcocystis infection in cattle in Qena Governorate, Upper Egypt both morphologically and molecularly. Sarcocystis cruzi and S. hjorti species were isolated from cattle, which is of veterinary importance and indicates that morphologically similar Sarcocystis species are genetically distinct. Additionally, the results show that Sarcocystis species are not host-specific.
| S-EPMC5825376 | biostudies-literature
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.
| S-EPMC9109842 | biostudies-literature
Project description:Mycoplasma pneumoniae is a major human respiratory pathogen causing both upper and lower respiratory disease in humans of all ages, and it can also result in other serious extrapulmonary sequelae. A multilocus sequence typing (MLST) scheme for M. pneumoniae was developed based on the sequences of eight housekeeping genes (ppa, pgm, gyrB, gmk, glyA, atpA, arcC, and adk) and applied to 55 M. pneumoniae clinical isolates and the two type strains M129 and FH. A total of 12 sequence types (STs) resulted for 57 M. pneumoniae isolates tested, with a discriminatory index of 0.21 STs per isolate. The MLST loci used in this scheme were shown to be stable in 10 strains following 10 sequential subculture passages. Phylogenetic analysis of concatenated sequences of the eight loci indicated two distinct genetic clusters that were directly linked to multilocus variable-number tandem repeat analysis (MLVA) type. Genetic MLST clustering was confirmed by genomic sequence analysis, indicating that the MLST scheme developed in this study is representative of the genome. Furthermore, this MLST scheme was shown to be more discriminatory than both MLVA and P1 typing for the M. pneumoniae isolates examined, providing a method for further and more detailed analysis of observed epidemic peaks of M. pneumoniae infection. This scheme is supported by a public Web-based database (http://pubmlst.org/mpneumoniae).
| S-EPMC4572520 | biostudies-literature
Project description:In this study we isolate and identify the Enteropathogenic Escherichia coli (EPEC) causing diarrhea in children less than five years in Cairo, Egypt, during different seasons. Children younger than five years with diarrhea, attending the Pediatric Gastroenterology Intensive Care Unit of the Cairo University Pediatric Hospital in one year period were our group of study. Our control group was age and sex matched concurrent healthy children. The identified E. coli isolates were subjected to antimicrobial disc diffusion susceptibility test and further identified for EPEC serotype by slide agglutination test, using antiserum E. coli somatic trivalent I (O111, O55, O26) according to the instructions of the manufacturer. Out of 134 patients 5.2% of them revealed EPEC in the fecal sample, while the 20 children control group showed no EPEC isolates in their samples. Our EPEC frequency showed variations from the compared results of other studies. Higher rate of EPEC (18.7%) was found in patients between 2 to 3 years, while EPEC rate was (7.5%) in patients less than 6 months old, with P < 0.05. EPEC was identified from fecal specimens as a unique pathogen or associated with other pathogens in acute and chronic diarrhea in children. EPEC were detected in all seasons except in winter, and was predominant in summer season. Four (57%) EPEC isolates were resistant to ampicillin, ticarcillin, and cotrimoxazole, and (14.3%) to the third generation cephalosporins.
| S-EPMC3254012 | biostudies-other
Project description:A total of 134 Egyptian liver flukes were collected from different definitive hosts (cattle, sheep, and buffaloes) to identify them via the use of PCR-RFLP and sequence analysis of the first nuclear ribosomal internal transcribed spacer (ITS1). Specimens of F. hepatica from France, as well as F. gigantica from Cameroon were included in the study for comparison. PCR products of ITS1 were subjected for digestion by RsaI restriction enzyme and visualized on agarose gel. According to RFLP pattern, Egyptian flukes were allocated into two categories. The first was identical to that of French hepatica flukes to have a pattern of 360, 100, and 60 (bp) band size, whereas the second resembled to that of Cameroonian gigantica worms to have a profile of 360, 170, and 60 bp in size. Results of RFLP analysis were confirmed by sequence analysis of representative ITS1 amplicons. No hybrid forms were detected in the present study. Taken together, this study concluded that both species of Fasciola are present in Egypt, whereas the hybrid form may be not very common.
| S-EPMC3671433 | biostudies-literature
Project description:Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) represent the most important avian Mycoplasma species in the poultry industry, causing considerable economic losses. In Italy, the presence of MG or MS has been investigated especially in commercial poultry farms. To our knowledge, no systematic investigations on MG or MS presence using highly specific diagnostic assays have been performed in backyard poultry. The aim of this study was to detect and molecularly characterize MG and MS strains in 11 backyard poultry flocks located in different regions of Italy. Tracheal swabs were collected and DNA was extracted. For MS, a PCR targeting a vlhA gene fragment was performed, and typing and subtyping was attempted. The presence of MG was investigated by a screening PCR, then MG typing by gene-targeted sequencing (GTS). All the amplicons were sequenced, then MG and MS dendrograms were constructed. All the flocks examined resulted Mycoplasma positive: 5 out of 11 (45.45%) were MG and MS positive, 3 (27.27%) were MG positive, and the remaining 3 (27.27%) were MS positive. The MS detections were assigned to types C, D, and F. All strains of type D belonged to subtype D1 and 2 unknown subtypes were identified. A MS sequence showed peculiar characteristics, which did not allow assignment to a known MS type or subtype. MG GTS analysis identified 6 MG strains belonging to 5 subclusters circulating in Italian backyards chicken flocks. The results of this study provide evidence of a risk for commercial poultry farms, especially in areas where backyard and commercial farms are close, suggesting the implementation of biosecurity measures.
| S-EPMC7587864 | biostudies-literature
Project description:Mycoplasma gallisepticum is the most virulent and economically important Mycoplasma species for poultry worldwide. Currently, M. gallisepticum strain differentiation based on sequence analysis of 5 loci remains insufficient for accurate outbreak investigation. Recently, whole-genome sequences (WGS) of many human and animal pathogens have been successfully used for microbial outbreak investigations. However, the massive sequence data and the diverse properties of different genes within bacterial genomes results in a lack of standard reproducible methods for comparisons among M. gallisepticum whole genomes. Here, we proposed the development of a core genome multilocus sequence typing (cgMLST) scheme for M. gallisepticum strains and field isolates. For development of this scheme, a diverse collection of 37 M. gallisepticum genomes was used to identify cgMLST targets. A total of 425 M. gallisepticum conserved genes (49.85% of M. gallisepticum genome) were selected as core genome targets. A total of 81 M. gallisepticum genomes from 5 countries on 4 continents were typed using M. gallisepticum cgMLST. Analyses of phylogenetic trees generated by cgMLST displayed a high degree of agreement with geographical and temporal information. Moreover, the high discriminatory power of cgMLST allowed differentiation between M. gallisepticum strains of the same outbreak. M. gallisepticum cgMLST represents a standardized, accurate, highly discriminatory, and reproducible method for differentiation among M. gallisepticum isolates. cgMLST provides stable and expandable nomenclature, allowing for comparison and sharing of typing results among laboratories worldwide. cgMLST offers an opportunity to harness the tremendous power of next-generation sequencing technology in applied avian mycoplasma epidemiology at both local and global levels.
| S-EPMC5744223 | biostudies-literature