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

2019 ◽  
Vol 12 (5) ◽  
pp. 664-670 ◽  
Author(s):  
Mounier M. Abdel Halium ◽  
Fayez A. Salib ◽  
S. A. Marouf ◽  
Emil S. Abdel Massieh
Keyword(s):  
Phylogenetic Analysis ◽  
Small Ruminants ◽  
Mycoplasma Species ◽  
African Countries ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Sheep And Goats ◽  
Pcr Product ◽  
Nasal Swabs ◽  
Bronchial Wash

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.

Molecular Detection of Bacterial Pathogens Directly from the Nasal Swabs and Lung Tissue of Sheep and Goats

2019 ◽  
Vol 15 (02) ◽  
pp. 22-25
Author(s):  
Sunaina Thakur ◽  
Subhash Verma ◽  
Prasenjit Dhar ◽  
Mandeep Sharma
Keyword(s):  
Pasteurella Multocida ◽  
Direct Detection ◽  
Bacterial Species ◽  
Economic Losses ◽  
Isolation And Identification ◽  
Sheep And Goats ◽  
Histophilus Somni ◽  
Nasal Swabs ◽  
Mycoplasma Spp

Respiratory infections of sheep and goats cause heavy morbidity and mortality, leading to huge economic losses. Conventional methods of diagnosis that include isolation and identification of incriminating microbes are time-consuming and fraught with logistic challenges. Direct detection of incriminating microbes using molecular tools is gaining popularity in clinical, microbiological settings. In this study, a total of 50 samples (44 nasal swabs and 6 lung tissues) from sheep and goats were screened for the detection of different bacterial species by in vitro amplification of genus or species-specific genes. Histophilus somni was detected in 2% goat samples, Trueperella pyogenes in 20% goat nasal swabs, whereas 22% goat nasal swab samples were found positive for Mycoplasma spp. None of the samples from sheep was detected positive for H. somni, T. pyogenes, Mycoplasma spp. Similarly, all samples, irrespective, whether from sheep or goats, showed negative results for Pasteurella multocida, Mannheimia haemolytica, and Corynebacterium pseudotuberculosis.

scholarly journals Comparison of nucleotide sequences of recent and previous lineages of peste-des-petits-ruminants viruses of sheep and goats in Nigeria

2016 ◽  
Vol 83 (1) ◽  
Author(s):  
Samuel Mantip ◽  
Melvyn Quan ◽  
David Shamaki ◽  
Moritz Van Vuuren
Keyword(s):  
Phylogenetic Analysis ◽  
Small Ruminants ◽  
Viral Disease ◽  
N Gene ◽  
Peste Des Petits Ruminants ◽  
East African ◽  
Tissue Samples ◽  
Sheep And Goats ◽  
Ecological Zones ◽  
Agro Ecological Zones

Peste-des-petits-ruminants virus (PPRV) is a highly contagious, fatal and economically important viral disease of small ruminants that is still endemic and militates against the production of sheep and goats in endemic areas of the world. The aim of this study was to describe the viral strains within the country. This was carried out by collecting tissue and swab samples from sheep and goats in various agro-ecological zones of Nigeria. The phylogeny of archived PPRV strains or isolates and those circulating and causing recent outbreaks was determined by sequencing of the nucleoprotein (N)-gene. Twenty tissue and swab samples from apparently healthy and sick sheep and goats were collected randomly from 18 states, namely 3 states in each of the 6 agro-ecological zones visited. A total of 360 samples were collected. A total of 35 samples of 360 (9.7%) tested positive by reverse transcriptase–polymerase chain reaction, of which 25 were from oculo-nasal swabs and 10 were from tissue samples. Neighbour-joining phylogenetic analysis using Phylogenetic Analysis Using Parsimony (PAUP) identified four different lineages, that is, lineages I, II, III and IV. Interestingly, the Nigerian strains described in this study grouped in two separate major lineages, that is, lineages II and IV. Strains from Sokoto, Oyo, Plateau and Ondo states grouped according to the historical distribution of PPRV together with the Nigerian 75/1 strain of lineage II, while other strains from Sokoto, Oyo, Plateau, Akwa-Ibom, Adamawa, Kaduna, Lagos, Bauchi, Niger and Kano states grouped together with the East African and Asian strains of lineage IV. This finding confirms that both lineage II and IV strains of PPRV are circulating in Nigeria. Previously, only strains of lineage II were found to be present in the country.

scholarly journals Isolation and identification of pseudomonas aeruginosa from infected sheep and detection of phosolipase C (lecithinase

2015 ◽  
Vol 39 (1) ◽  
pp. 28-32
Author(s):  
Ahmed N. Mahmood
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Egg Yolk ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Morphological Examination ◽  
Isolation And Identification ◽  
Nasal Swabs ◽  
Green Zone ◽  
Eye Infection ◽  
Agar Well Diffusion Method

The current study dealt with the isolation and identification of 19 isolates (14.07%) for the period (October 2013 to January 2014) from different samples (135 samples) number of isolates belonging to Pseudomonas aeruginosa were 8 isolated from urine samples, their percentage were 33.33 % and 5 isolates from nasal swabs, (11.11%) and 4 isolated from milk samples (9.30%), 2 isolated from wound swabs (24%) and isolates from eye infection (9.09%). These isolates were identified by morphological examination and biochemical tests and API-20 NE system. The second part was the study of the antibiotic susceptibility that was carried out on 19 isolates of P. aeruginosa for 12 types of antibiotic. The results showed that isolates were resistant to 9 out of 12 antibiotics with percentage (Penicillin 100%, nalidixic acid100%, pipracillin 100%, erythromycin 68,4%, Chloramphenicol 63%, 78%, trimthoprim 73%, ceftacizidem 89%, cefitriaxon 84% and cefotaxime 89%) on the other hand most of isolates were sensitive to (epipenime, azythromycin, and polymixin). Detection of phospholipase C (Lecithinase) by agar well diffusion method on different selective media as (egg yolk agar, briliant green crystal violates lecithnase agar BCL), 12 isolates out of 19 showed a positive reaction on these media. The result depended on measurement the diameter of opacity zone produced and the blush green zone on the BCL media, the diameter of zone ranged from 8mm to 32mm. both media (EYA and BCL media is the best media for detection of phospholipase by P. aeruginosa.

scholarly journals Comparison of Coxiella burnetii Excretion between Sheep and Goats Naturally Infected with One Cattle-Associated Genotype

Pathogens ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 652
Author(s):  
Benjamin Bauer ◽  
Louise Prüfer ◽  
Mathias Walter ◽  
Isabel Ganter ◽  
Dimitrios Frangoulidis ◽  
...  
Keyword(s):  
Public Health ◽  
Environmental Samples ◽  
Small Ruminants ◽  
Infected Sheep ◽  
Whole Genome ◽  
Typing Method ◽  
Sheep And Goats ◽  
Rectal Route ◽  
Nasal Swabs ◽  
Species Specific

The main reservoir of Coxiella (C.) burnetii are ruminants. They shed the pathogen through birth products, vaginal mucus, faeces and milk. A direct comparison of C. burnetii excretions between naturally infected sheep and goats was performed on the same farm to investigate species-specific differences. The animals were vaccinated with an inactivated C. burnetii phase I vaccine at the beginning of the study period for public health reasons. Vaginal and rectal swabs along with milk specimens were taken monthly during the lambing period and once again at the next lambing season. To estimate the environmental contamination of the animals’ housings, nasal swabs from every animal were taken simultaneously. Moreover, dust samples from the windowsills and straw beddings were collected. All samples were examined by qPCR targeting the IS1111 gene and the MLVA/VNTR typing method was performed. Whole genome sequencing was applied to determine the number of IS1111 copies followed by a calculation of C. burnetii genome equivalents of each sample. The cattle-associated genotype C7 was detected containing 29 IS1111 copies. Overall, goats seem to shed more C. burnetii through vaginal mucus and in particular shed more and for longer via the rectal route than sheep. This is supported by the larger quantities of C. burnetii DNA detected in caprine nasal swabs and environmental samples compared to the ovine ones. Transmission of C. burnetii from cattle to small ruminants must also be considered.

scholarly journals Evaluation of Risk Factors for Peste des Petits Ruminants Virus in Sheep and Goats at the Wildlife-Livestock Interface in Punjab Province, Pakistan

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Aziz-ul-Rahman ◽  
Muhammad Abubakar ◽  
Muhammad Hidayat Rasool ◽  
Shumaila Manzoor ◽  
Muhammad Saqalein ◽  
...  
Keyword(s):  
Risk Factors ◽  
Significant Risk ◽  
Small Ruminants ◽  
Significant Risk Factor ◽  
N Gene ◽  
High Morbidity ◽  
Peste Des Petits Ruminants ◽  
Sheep And Goats ◽  
Wild Ruminants ◽  
Nasal Swabs

Peste des petits ruminants virus (PPRV) is causing infectious disease with high morbidity and mortality rate in domestic and wild small ruminants of Pakistan with valuable economical losses. The present study was carried out to investigate risk factors of PPRV in domestic small ruminants which were present in the vicinity of wildlife parks. A total of 265 sera samples (27 wild ruminants and 238 domesticated small ruminants) from apparently healthy animals from two different wildlife parks were collected and analysed for PPRV antibodies. Also, 20 nasal swabs from domestic small ruminants showing respiratory signs were collected to check for presence of PPRV antigen. Competitive ELISA revealed highest proportions of anti-PPRV antibodies in domestic small ruminants around the Wildlife Park at Lahore (35%) as compared to Faisalabad (13%), with no existence of PPRV antibodies in tested serum of wild ruminants at these parks. Higher seropositivity was observed in females (25.6%) than in males (5.1%) and in goats (34.5%) compared to sheep (11.2%). The results of N-gene based RT-PCR highlight the absence of PPRV due to lack of current PPR outbreak in the region during study period. Even though grazing was not a significant risk factor, there is still a possibility of wildlife-livestock interactions for feed and water reservoirs, resulting in spillover of PPR to wildlife. Keeping in view the high seropositivity and risk of PPR, vaccination should be adopted to avoid circulation of PPRV among wild and domestic small ruminants (sheep and goats).

scholarly journals Isolation and Identification of Bacteria from the Workers of Live Bird Markets at Mymensingh, Bangladesh

1970 ◽  
Vol 17 ◽  
pp. 135-138
Author(s):  
S Sarker ◽  
S Talukder ◽  
EH Chowdhury ◽  
PM Das
Keyword(s):  
Escherichia Coli ◽  
Dna Isolation ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
E Coli ◽  
Live Bird Markets ◽  
Nasal Swabs ◽  
Identification Of Bacteria ◽  
Live Bird

Context: Identification of bacteria from the workers of live bird markets is important factor for zoonotic aspects and for implementing appropriate control strategies.Objectives: To determine the occurrence of bacteria especially Salmonella sp. and Escherichia coli from the workers of live bird markets.Materials and Methods: A total of 40 samples were collected from hand washes (n=20) and nasal swabs (n=20) of the associated workers in urban and suburban live bird markets. Bacteria were isolated in different media, and identification was performed based on the staining, cultural and some biochemical tests. For Salmonella sp., DNA was extracted using a DNA isolation kit and rfbs gene was amplified by using commercial PCR kit.Results: The bacteria such as Salmonella sp. and E. coli were detected in the samples by several microbial tests. The prevalence of Salmonella sp. was 40% and 30%, and E. coli was 70% and 40% in the hand washes and nasal swabs respectively of the workers of urban and periurban live bird markets.Conclusion: The results obtained in this study suggest that the appropriate precautions should be taken during and subsequent to the handling of live birds to minimize the risk of zoonotic diseases.Key words: Salmonella; Escherichia coli; live bird markets; isolation and identificationDOI: 10.3329/jbs.v17i0.7121J. bio-sci. 17: 135-138, 2009

scholarly journals Clinical, histopathological, and molecular characterization of Mycoplasma species in sheep and goats in Egypt

2021 ◽  
pp. 2561-2567
Author(s):  
Walid S. Mousa ◽  
Ahmed A. Zaghawa ◽  
Ahmed M. Elsify ◽  
Mohamed A. Nayel ◽  
Zarroug H. Ibrahim ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Respiratory Infections ◽  
Histopathological Examination ◽  
Small Ruminants ◽  
Systemic Reaction ◽  
Economic Losses ◽  
Mycoplasma Species ◽  
Nasal Discharge ◽  
Sheep And Goats

Background and Aim: Mycoplasma infection in small ruminants is a serious problem in sheep and goat herds around the world. It is responsible for high economic losses and decreased animal productivity. This study aimed to highlight the clinical, histopathological, minimum inhibitory concentration (MIC), and molecular characterization of Mycoplasma species in sheep and goats in Menoufiya Governorate, Egypt. Materials and Methods: A total of 234 samples were collected; 104 samples were collected from pneumonic lung tissues from the abattoir, in addition, 10 and 20 samples collected from apparently and diseased sheep, respectively, and 40 and 60 samples were collected from apparently and diseased goats, respectively, which were subjected to isolation onto pleuropneumonia-like organism medium. Polymerase chain reaction (PCR), histopathological examination, and determination of the MIC were also performed. Results: Of 104 samples of lung tissues showing pneumonic lesions, 56 (53.84%) were positive for Mycoplasma isolation. The positive isolation of Mycoplasma from 10 and 20 samples from apparently and diseased sheep was 30% and 40%, respectively as well as the positive isolation of Mycoplasma was 17% and 56.66% out of 40 and 60 apparently healthy and diseased field goat's cases, respectively. All the diseased sheep and goats showed respiratory manifestations, including cough, bilateral nasal discharge, conjunctivitis, and systemic reaction. Evaluation of the MIC for Mycoplasma ovipneumoniae revealed that lincospectin and tylosin were the most effective antibiotics at 2.5 μg/mL. Histopathological examination of affected lung tissue showed extensive hemorrhagic pneumonia with extensive alveolar hemorrhage. The PCR technique proved to be a rapid, specific, and sensitive method for the detection of M. ovipneumoniae and Mycoplasma arginini at 390 and 326 bp, respectively. Conclusion: M. ovipneumoniae and M. arginini were the most prevalent species associated with respiratory infections in sheep and goats in the study area. Further studies are needed to investigate the role of these species in dissemination of the disease within herds of small ruminants.

scholarly journals Gene Sequencing and Phylogenetic Analysis: Powerful Tools for an Improved Diagnosis of Fish Mycobacteriosis Caused by Mycobacterium fortuitum Group Members

2021 ◽  
Vol 9 (4) ◽  
pp. 797
Author(s):  
Davide Mugetti ◽  
Mattia Tomasoni ◽  
Paolo Pastorino ◽  
Giuseppe Esposito ◽  
Vasco Menconi ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Gene Sequencing ◽  
Diagnostic Techniques ◽  
Individual Species ◽  
Identification System ◽  
Mycobacterium Fortuitum ◽  
Biochemical Tests ◽  
Species Determination ◽  
Gene Encoding ◽  
The Individual

The Mycobacterium fortuitum group (MFG) consists of about 15 species of fast-growing nontuberculous mycobacteria (NTM). These globally distributed microorganisms can cause diseases in humans and animals, especially fish. The increase in the number of species belonging to MFG and the diagnostic techniques panel do not allow to clarify their real clinical significance. In this study, biomolecular techniques were adopted for species determination of 130 isolates derived from fish initially identified through biochemical tests as NTM belonging to MFG. Specifically, gene sequencing and phylogenetic analysis were used based on a fragment of the gene encoding the 65 KDa heat shock protein (hsp65). The analyzes made it possible to confirm that all the isolates belong to MFG, allowing to identify the strains at species level. Phylogenetic analysis substantially confirmed what was obtained by gene sequencing, except for six strains; this is probably due to the sequences present in NCBI database. Although the methodology used cannot represent a univocal identification system, this study has allowed us to evaluate its effectiveness as regards the species of MFG. Future studies will be necessary to apply these methods with other gene fragments and to clarify the real pathogenic significance of the individual species of this group of microorganisms.

scholarly journals Molecular Detection and Phylogeny of Anaplasmaphagocytophilum and Related Variants in Small Ruminants from Turkey

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 814
Author(s):  
Münir Aktaş ◽  
Sezayi Özübek ◽  
Mehmet Can Uluçeşme
Keyword(s):  
16S Rrna ◽  
Small Ruminants ◽  
Rrna Gene ◽  
Infection Rates ◽  
Sheep And Goats ◽  
Significant Difference ◽  
Pcr Rflp ◽  
First Time ◽  
Japan And China ◽  
Apparently Healthy

Anaplasma phagocytophilum causes tick-borne fever in small ruminants. Recently, novel Anaplasma variants related to A. phagocytophilum have been reported in ruminants from Tunisia, Italy, South Korea, Japan, and China. Based on 16S rRNA and groEL genes and sequencing, we screened the frequency of A. phagocytophilum and related variants in 433 apparently healthy small ruminants in Turkey. Anaplasma spp. overall infection rates were 27.9% (121/433 analyzed samples). The frequency of A. phagocytophilum and A. phagocytophilum-like 1 infections was 1.4% and 26.5%, respectively. No A. phagocytophilum-like 2 was detected in the tested animals. The prevalence of Anaplasma spp. was comparable in species, and no significant difference was detected between sheep and goats, whereas the prevalence significantly increased with tick infestation. Sequencing confirmed PCR-RFLP data and showed the presence of A. phagocytophilum and A. phagocytophilum-like-1 variant in the sampled animals. Phylogeny-based on 16S rRNA gene revealed the A. phagocytophilum-like 1 in a separate clade together with the previous isolates detected in small ruminants and ticks. In this work, A. phagocytophilum-like 1 has been detected for the first time in sheep and goats from Turkey. This finding revealed that the variant should be considered in the diagnosis of caprine and ovine anaplasmosis.

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