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Abstract This study addresses the distribution and antimicrobial resistance of Acinetobacter baumannii (A. baumannii) in a medical facility in Haikou City, aiming to provide essential insights for enhancing in-hospital treatment and prevention strategies. We conducted a retrospective analysis of 513 A. baumannii isolates collected from a tertiary care hospital in Haikou between January 2018 and December 2020, focusing on their antimicrobial resistance patterns. Random Amplified Polymorphic DNA (RAPD) analysis was performed on 48 randomly selected A. baumannii strains. Using Gel-pro4.0 and NTSYSspc2.10 software, we constructed dendrograms to assess the genetic diversity of these strains. Our results indicate that males between 60 and 70 years old are particularly vulnerable to A. baumannii infections, which are most frequently detected in sputum samples, with a detection rate exceeding 70%. Alarmingly, over 50% of the isolates were identified as multi-drug resistant. The RAPD-PCR fingerprinting cluster analysis demonstrated substantial genetic diversity among the strains. Using primer OPA-02 at a 45% similarity coefficient, the strains were categorized into four groups (A-D), with group A being predominant (39 strains). high-prevalence areas like the Neurosurgery and Intensive Care Medicine Wards require enhanced surveillance and targeted interventions to manage Group C infections effectively. Additionally, the varied presence of other groups necessitates customized strategies to address the specific risks in each ward. Similarly, primer 270 at a 52% similarity coefficient classified the strains into five groups (E-I), with group E being most common (36 strains). The study highlights a concerning prevalence of antimicrobial resistance, particularly multi-drug resistance, among A. baumannii strains in the Haikou hospital. The significant genetic diversity, especially within groups A and E, underscores the need for tailored hospital treatment protocols and prevention measures. These findings contribute to the growing body of research on antimicrobial resistance, emphasizing the urgent need for effective management strategies in healthcare settings.

Abstract Background Trueperella pyogenes is an opportunistic pathogen that causes suppurative infections in various animal species, including goats. So far, only limited knowledge of phenotypic and genotypic properties of T. pyogenes isolates from goats has been gathered. In our study, we characterized the phenotypic and genotypic properties of caprine T. pyogenes isolates and established their relationship by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR). Results From 2015 to 2023, 104 T. pyogenes isolates were obtained from 1146 clinical materials. In addition, two T. pyogenes isolates were obtained from 306 swabs collected from healthy goats. A total of 51 T. pyogenes isolates were subjected to detailed phenotypic and genotypic characterization. The virulence genotype plo/nanH/nanP/fimA/fimC/luxS was predominant. All of the tested isolates showed the ability to form a biofilm but with different intensities, whereby most of them were classified as strong biofilm formers (72.5%). The high level of genetic diversity among tested caprine T. pyogenes isolates (19 different RAPD profiles) was observed. The same RAPD profiles were found for isolates obtained from one individual, as well as from other animals in the same herd, but also in various herds. Conclusions This study provided important data on the occurrence of T. pyogenes infections in goats. The assessment of virulence properties and genetic relationships of caprine T. pyogenes isolates contributed to the knowledge of the epidemiology of infections caused by this pathogen in small ruminants. Nevertheless, further investigations are warranted to clarify the routes of transmission and dissemination of the pathogen.

We evaluated the specificity of the primers OPF-01, P54, and 1253 to identify A. fumigatus, A. flavus, and A. niger, respectively, with the RAPD-PCR method. Eighty-two isolates belonging to the sections Fumigati, Flavi, and Nigri were used. The isolates were identified by phenotypic (macro- and micromorphology) and genotypic (partial sequences of the BenA gene) methods. The RAPD-PCR method was used to obtain polymorphic patterns with the primers OPF-01, P54, and 1253. The specificity of the polymorphic patterns of the isolates of each species was evaluated through the UPGMA clustering method and logistic regression model. All isolates of the genus Aspergillus were identified at the section level by macro- and micromorphology showing the typical morphology of the sections Fumigati, Flavi, and Nigri, and the species were identified by the construction of the phylogeny of the partial sequence of the BenA gene. The patterns' polymorphic strains obtained with the primers OPF-01, P54, and 1253 for the isolates of A. fumigatus, A. flavus, and A niger, respectively, showed the same polymorphic pattern as the reference strains for each species. To verify the specificity of the primers, they were tested with other species from the sections Fumigati, Flavi and Nigri. The results support that the primers OPF-01, P54, and 1253 generate polymorphic patterns by RAPD-PCR species specific to A. fumigatus, A. flavus, and A. niger, respectively. [ABSTRACT FROM AUTHOR]

Dermatophytes are the keratinophilic fungi which infect humans and is the most recurring type of disease. The high level of transmissibility creates an epidemiological risk and emphasises the significance of these illnesses. However, a growing number of reports describing dermatophytes can cause deep infections in diabetic and immunocompromised patients, by invading deep layers like the dermis and hypodermis. Despite the prevalence and significance of dermatophytes in clinical mycology, it is not always possible to accurately diagnose this specific infection due to its overlapping structures among species of dermatophytes. Since it is difficult to identify species that exhibit weak characteristics in the morphological highlights, identification of the dermatophyte is often relied on its morphological analysis, which is a laborious process and demands skill. The massive shift in genetic variation, the source of infection, and epidemiological research can be discovered using molecular approaches. Therefore, the development of an accurate laboratory test for dermatophyte species identification is essential for the prevention and efficient management of dermatophytoses. One such methodology allows use of PCR technology which has many methods for molecular level characterization which is rapid, efficient, and capable of producing DNA polymorphisms specific to various dermatophyte species based on distinctive band patterns seen by agarose gel electrophoresis. The RAPD-PCR approach will be used in this study protocol to molecularly characterize the dermatophytes for precise speciation of the sample. In addition to improving knowledge of fungal biology and pathology with a focus on adaptive mechanisms to combat difficult conditions from host counteractions, there is a need to improve awareness of the importance of these diseases through accurate epidemiological data. The advantages of molecular approaches for characterizing objects over traditional methods are their sensitivity and specificity.

Background: The main causes of hospital- and community-acquired urinary tract infections (UTIs) are a group of Escherichia coli (E. coli) strains with multiple virulence factors known as uropathogenic E. coli. Methods and results: One hundred E. coli isolates from the urine specimens of hospital- and community-acquired UTI patients were characterized based on their virulence factors and genetic relatedness using PCR and RAPD‒PCR, respectively. Among all, the traT (71%), sitA (64%), ompT (54%), malX (49%), ibeA (44%), tsh (39%), hlyD (18%) and cnf1 (12%) genes had the highest to lowest frequencies, respectively. There was no significant difference between the frequency of tested virulence genes in E. coli isolates from inpatients and outpatients. The frequency of the hlyD gene was significantly greater in E. coli isolates from patients hospitalized in gynecology, dermatology and intensive care unit (ICU) wards than in those from other wards. Eight virulence gene patterns were common among the isolates of inpatients in different wards of the same hospital, of which five patterns belonged to the isolates of inpatients in the same ward. More E. coli isolates with similar virulence gene patterns and greater genetic similarity were found in female patients than in male patients. The analysis of the RAPD‒PCR dendrograms revealed more genetic similarities among the E. coli isolates from inpatients than among those from outpatients. Conclusion: Our findings indicate the presence of a wide variety of virulence factors in E. coli isolates and the possibility of spreading the same clones in different wards of the hospital. [ABSTRACT FROM AUTHOR]

In this study, molecular typing using Randomly Amplified Polymorphic DNA (RAPD-PCR) was conducted on 16 original isolates of Metarhizium acridum obtained from locusts (Schistocerca piceifrons ssp. piceifrons.) in Mexico (MX). The analysis included reference strains of the genus Metarhizium sourced from various geographical regions. The isolates were identified by phenotypic (macro and micromorphology) and genotypic methods (RAPD-PCR and Amplified Fragment Length Polymorphisms (AFLP), through a multidimensional analysis of principal coordinates (PCoA) and a minimum spanning network (MST). Subsequently, Sequences-Characterized Amplified Region (SCAR) markers were developed for the molecular detection of M. acridum, these markers were chosen from polymorphic patterns obtained with 14 primers via RAPD-PCR. Phenotypic and genotypic characterization identified the MX isolates as M. acridum. Of all the polymorphic patterns obtained, only OPA04 and OPA05 were chosen, which presented species-specific bands for M. acridum, and further utilized to create SCAR markers through cloning and sequencing of the specific bands. The specificity of these two markers was confirmed via Southern hybridization. The SCAR markers (Ma-160OPA-05 and Ma-151OPA-04) exhibit remarkable sensitivity, detecting down to less than 0.1 ng, as well as high specificity, as evidenced by their inability to cross-amplify or generate amplification with DNAs from other strains of Metarhizium (as Metarhizium anisopliae) or different genera of entomopathogenic fungi (Cordyceps fumosorosea and Akanthomyces lecanii). These SCAR markers yield readily detectable results, showcasing high reproducibility. They serve as a valuable tool, especially in field applications. [ABSTRACT FROM AUTHOR]

Pollutants in an environment can have long-term implications for the species living there, resulting in local adaptations with implications for their genetic structure. Heavy metal pollutants infiltrate soils and groundwater, bioaccumulate in food webs, and negatively impact biota. In this study, we investigated the degree to which the genetic structure and variability of the slender green-winged grasshopper (Aiolopus thalassinus (Fabricius) (Orthoptera: Acrididae)) were impacted by heavy metal pollution and distance. We used the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method to examine the genetic variability of populations in 3 heavy metal-polluted and 3 unpolluted locations across varying geographical distances in Egypt. The heavy metal concentrations of cadmium, copper, lead, and zinc were measured from the grasshopper tissue and soils. Sixty-nine unique and polymorphic bands were produced by 4 primers. Cluster and principal component analyses separated the populations inside and outside Cairo into 2 main branches, which were further divided into smaller branches corresponding to their geographical regions. We found no differences in the Shannon genetic diversity index between populations or with increasing heavy metal concentrations in either the soil or the grasshopper tissue. Our results showed a greater genetic variation among populations than between populations within the same location, indicating populations within locations were less differentiated than those between locations. The moderate correlation between genetic similarity and spatial distance suggests geographical isolation influenced grasshopper population differentiation. Based on the RAPD analysis, environmental pollutants and geographical distances impact the A. thalassinus population structure, potentially restricting gene flow between sites even at small spatial scales. [ABSTRACT FROM AUTHOR]

Synthetic seeds are defined as any tissue with the potential to grow into a plant under in vitro or in vivo conditions that has been artificially encapsulated somatic embryos, shoot buds, cell aggregates, or other tissues. These tissues retain their potentiality even after being stored. Somatic embryos used in plant production and dispersed in the field were considered synthetic seeds. After using artificial seeds, shoot apical tips, axillary buds, and nodal segments have also been used as suitable substitutes for somatic embryos. The current study's goal was to standardize artificial seed production methods using Populus alba encapsulated nodal explants and in vitro regeneration. This woody species is male tree and has no seed formation. Several concentrations of sodium alginate (2, 3, 4 and 5%) were mixed separately with the somatic endosperm of P. alba; and finally encapsulated with various concentrations of CaCl2 (50, 75, 100 and 200mM). The morphological and physiological parameters were measured and sometimes showed significant differences. The best conditions for these synthetic seeds' formation based on the morphological and physiological parameters were the beads with 3 and 4% sodium alginate which are encapsulated by 100 and 200mM of CaCl2. The current study clearly established and demonstrated the best method for producing artificial seed in P. alba. Key message: Synthetic seeds are modern application for tissue culture to solve germination problem, either in non-germinated plants or poor-germinated ones. These seeds germinated with higher productivity comparing to plantlets from micropropagation. [ABSTRACT FROM AUTHOR]

Abstract Background Curbing the potential negative impact of antibiotic resistance, one of our era's growing global public health crises, requires regular monitoring of the resistance situations, including the reservoir of resistance genes. Wild birds, a possible bioindicator of antibiotic resistance, have been suggested to play a role in the dissemination of antibiotic-resistant bacteria. Therefore, this study was conducted with the objective of determining the phenotypic and genotypic antibiotic resistance profiles of 100 Escherichia coli isolates of gull and pigeon origin by using the Kirby-Bauer disk diffusion method and PCR. Furthermore, the genetic relationships of the isolates were determined by RAPD-PCR. Results Phenotypic antibiotic susceptibility testing revealed that 63% (63/100) and 29% (29/100) of E. coli isolates were resistant to at least one antibiotic and multidrug-resistant (MDR), respectively. With the exception of cephalothin, to which the E. coli isolates were 100% susceptible, tetracycline (52%), kanamycin (38%), streptomycin (37%), ampicillin (28%), chloramphenicol (21%), trimethoprim/sulfamethoxazole (19%), gentamicin (13%), enrofloxacin (12%) and ciprofloxacin (12%) resistances were detected at varying degrees. Among the investigated resistance genes, tet(B) (66%), tet(A) (63%), aphA1 (48%), sul3 (34%), sul2 (26%), strA/strB (24%) and sul1 (16%) were detected. Regarding the genetic diversity of the isolates, the RAPD-PCR-based dendrograms divided both pigeon and gull isolates into five different clusters based on a 70% similarity threshold. Dendrogram analysis revealed 47–100% similarities among pigeon-origin strains and 40–100% similarities among gull-origin E.coli strains. Conclusions This study revealed that gulls and pigeons carry MDR E. coli isolates, which may pose a risk to animal and human health by contaminating the environment with their feces. However, a large-scale epidemiological study investigating the genetic relationship of the strains from a "one health" point of view is warranted to determine the possible transmission patterns of antibiotic-resistant bacteria between wild birds, the environment, humans, and other hosts. Video Abstract

The investigation of genetic differentiation of subspecies and breeds is nowadays fundamental for understanding their adaptation to specific environments and for facilitating selection processes for production purposes. These aspects allow us both to safeguard their biodiversity and to protect the products with a high nutritional value that derive from them and to which consumers today pay particular importance.In this study, a genomic approach by RAPD-PCR and DNA-barcoding was used to identify possible genetic markers to discriminate Italian Mediterranean Buffalo (IMB) and Romanian Buffalo (RB). The two molecular tools have been widely used in this study not only to clarify genetic differences between breeds but also to avoid food frauds, given the great importance of Italian buffalo mozzarella recognized all over the world and protected by Denomination of Protected Origin (DPO).The research did not reveal any difference between the two breeds, despite belonging to two geographically different areas.

Dermatophytes are the keratinophilic fungi which infect humans and is the most recurring type of disease. The high level of transmissibility creates an epidemiological risk and emphasises the significance of these illnesses. However, a growing number of reports describing dermatophytes can cause deep infections in diabetic and immunocompromised patients, by invading deep layers like the dermis and hypodermis. Despite the prevalence and significance of dermatophytes in clinical mycology, it is not always possible to accurately diagnose this specific infection due to its overlapping structures among species of dermatophytes. Since it is difficult to identify species that exhibit weak characteristics in the morphological highlights, identification of the dermatophyte is often relied on its morphological analysis, which is a laborious process and demands skill. The massive shift in genetic variation, the source of infection, and epidemiological research can be discovered using molecular approaches. Therefore, the development of an accurate laboratory test for dermatophyte species identification is essential for the prevention and efficient management of dermatophytoses. One such methodology allows use of PCR technology which has many methods for molecular level characterization which is rapid, efficient, and capable of producing DNA polymorphisms specific to various dermatophyte species based on distinctive band patterns seen by agarose gel electrophoresis. The RAPD-PCR approach will be used in this study protocol to molecularly characterize the dermatophytes for precise speciation of the sample. In addition to improving knowledge of fungal biology and pathology with a focus on adaptive mechanisms to combat difficult conditions from host counteractions, there is a need to improve awareness of the importance of these diseases through accurate epidemiological data. The advantages of molecular approaches for characterizing objects over traditional methods are their sensitivity and specificity.

Background: Curbing the potential negative impact of antibiotic resistance, one of our era's growing global public health crises, requires regular monitoring of the resistance situations, including the reservoir of resistance genes. Wild birds, a possible bioindicator of antibiotic resistance, have been suggested to play a role in the dissemination of antibiotic-resistant bacteria. Therefore, this study was conducted with the objective of determining the phenotypic and genotypic antibiotic resistance profiles of 100 Escherichia coli isolates of gull and pigeon origin by using the Kirby-Bauer disk diffusion method and PCR. Furthermore, the genetic relationships of the isolates were determined by RAPD-PCR. Results: Phenotypic antibiotic susceptibility testing revealed that 63% (63/100) and 29% (29/100) of E. coli isolates were resistant to at least one antibiotic and multidrug-resistant (MDR), respectively. With the exception of cephalothin, to which the E. coli isolates were 100% susceptible, tetracycline (52%), kanamycin (38%), streptomycin (37%), ampicillin (28%), chloramphenicol (21%), trimethoprim/sulfamethoxazole (19%), gentamicin (13%), enrofloxacin (12%) and ciprofloxacin (12%) resistances were detected at varying degrees. Among the investigated resistance genes, tet(B) (66%), tet(A) (63%), aphA1 (48%), sul3 (34%), sul2 (26%), strA/strB (24%) and sul1 (16%) were detected. Regarding the genetic diversity of the isolates, the RAPD-PCR-based dendrograms divided both pigeon and gull isolates into five different clusters based on a 70% similarity threshold. Dendrogram analysis revealed 47–100% similarities among pigeon-origin strains and 40–100% similarities among gull-origin E.coli strains. Conclusions: This study revealed that gulls and pigeons carry MDR E. coli isolates, which may pose a risk to animal and human health by contaminating the environment with their feces. However, a large-scale epidemiological study investigating the genetic relationship of the strains from a "one health" point of view is warranted to determine the possible transmission patterns of antibiotic-resistant bacteria between wild birds, the environment, humans, and other hosts. 3bRmtrrdv9vbDYjBBmhJ2h Video Abstract [ABSTRACT FROM AUTHOR]

Purpose: The current study aims to investigate genotypic and phenotypic aspects of methicillinresistant Staphylococcus epidermidis (MRSE) strains involved in the biofilm formation and the attendance of the icaADB gene and genotypic characterization of this gene by random amplified polymorphic DNA (RAPD) PCR. Methods: 60 Staphylococcus epidermidis strains were isolated from clinical specimens, suspected of having the bacteria, from the laboratories of Isfahan. Biofilm formation was measured by the microtiter plate method. The attendance of biofilm formation genes was studied using PCR and all isolates producing biofilm (strong and moderate) were genetically classified by RAPD-PCR. Results: 37 isolates (61.7%) were MRSE and all positive biofilm strains. The prevalence of biofilm-related genes in the isolates was SesC (100%), SesI (45.9%), icaA (29.7%), icaB (37.8%), icaC (81.08%), icaD (70.2%), arcA (81.08%), and opp3AB (70%). PCR analysis showed that among 30 isolates of strong and medium biofilm production, 70% (21/30) positive for the icaADB gene. The Dendrograms obtained from RAPD-PCR results showed that all nine main clusters were at an 80% similarity level, and there were four isolates of a single type. Conclusion: These findings confirmed the high genotypic diversity of biofilm-producing MRSE strains and the relative diffusion of specific clones among clinical samples. [ABSTRACT FROM AUTHOR]

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Background: Food safety is an important subject that the global cheese industry increases awareness of. This urges these economic sectors to elevate the level of research to minimize cheese contamination with pathogenic bacteria, such as Salmonella. Aim: Based on these merits, this study was conducted to genotype Salmonella spp. isolated from cheese samples of local stores in Al-Diwaniyah City, Iraq. Methods: The study used 41 samples of local fresh unsalted white cheese in a selective-growth-based isolation of Salmonella. These isolates were confirmed utilizing a slide-agglutination (SA) test and VITEK® 2 system (V2S). Then, the isolates were subjected to conventional PCR and sequencing techniques that both targeted the 16S rRNA gene. For subtyping, the Salmonella isolates were subjected to a random amplified polymorphic DNA (RAPD)-PCR method. Results: The results of both SA and V2S revealed the presence of 14 (34.2%) isolates of Salmonella spp. in the cheese samples. The PCR confirmed 6 (42.9%) of these isolates, which further were defined with close nucleotide similarity (98.03%) and (97.88%) to different world isolates, such as Salmonella enterica subsp. Arizonae and Salmonella enterica subsp. enterica serovar Typhi, respectively. The RAPD-PCR findings showed different fragments for all the tested isolates. Conclusion: The present study indicates that the samples of the local fresh unsalted white cheese contain different Salmonella genotypes, which could be originated from different contamination sources. [ABSTRACT FROM AUTHOR]

The investigation of genetic differentiation of subspecies and breeds is nowadays fundamental for understanding their adaptation to specific environments and for facilitating selection processes for production purposes. These aspects allow us both to safeguard their biodiversity and to protect the products with a high nutritional value that derive from them and to which consumers today pay particular importance. In this study, a genomic approach by RAPD-PCR and DNA-barcoding was used to identify possible genetic markers to discriminate Italian Mediterranean Buffalo (IMB) and Romanian Buffalo (RB). The two molecular tools have been widely used in this study not only to clarify genetic differences between breeds but also to avoid food frauds, given the great importance of Italian buffalo mozzarella recognized all over the world and protected by Denomination of Protected Origin (DPO). The research did not reveal any difference between the two breeds, despite belonging to two geographically different areas. [ABSTRACT FROM AUTHOR]

Background: Food safety is an important subject that global cheese industry increases the awareness upon. This urges these economic sectors to elevate the level of research to minimize cheese contamination with pathogenic bacteria, such as Salmonella. Aim: Based on these merits, this study was conducted to genotype Salmonella spp isolated from cheese samples of local stores in Al-Diwaniyah City, Iraq. Methods: The study used 41 samples of local fresh unsalted white cheese in a selective-growth based isolation of Salmonella. These isolates were confirmed utilizing a slide-agglutination (SA) test and VITEK® 2 system (V2S). Then, the isolates were subjected to conventional PCR and sequencing techniques that both targeted the 16S rRNA gene. For subtyping, the Salmonella isolates were subjected to a random amplified polymorphic DNA (RAPD)-PCR method. Results: The results of both SA and V2S revealed the presence of 14 (34.2%) isolates of Salmonella spp. in the cheese samples. The PCR confirmed 6 (42.9%) of these isolates, which further were defined with close nucleotide similarity (98.03%) and (97.88%) to different world isolates, such as Salmonella enterica subsp. Arizonae and Salmonella enterica subsp. enterica serovar Typhi, respectively. The RAPD-PCR findings showed different fragments for all the tested isolates. Conclusion: The present study indicates that the samples of the local fresh unsalted white cheese contain different Salmonella genotypes, which could be originated from different contamination sources. [Open Vet J 2023; 13(10.000): 1277-1282]

Background: Healthcare-associated infections appear in a patient receiving medical attention at a hospital or other healthcare facility and develops infections that weren't present when they were admitted. One of the most important organisms causing infections acquired in hospitals is Staphylococcus aureus (S. aureus). For many bacterial species, Random Amplified Polymorphic DNA (RAPD) is a low-cost but effective typing technique. Objectives: To determine the genotypic polymorphism, the level of genetic relatedness, and the antimicrobial sensitivity and resistance among various S. aureus isolates. Methods: The following tests were performed on 50 samples that were obtained using strict aseptic precautions: sample culture, isolate identification using colony morphology, Gram stained film, biochemical responses, and antibiotic susceptibility testing. After RAPD-PCR, Syngene gene tool software was used to analyse the fingerprint pattern. Results: The most prevalent isolated organism was S. aureus. as it represented (70%) of isolated pathogens. Vancomycin and linezolid were effective against S. aureus isolates. The isolates of S. aureus were more common in nasal swab (42.9%) followed by wound (28.5%) then urine &endotracheal aspirate (14.3%). Patterns of RAPD-PCR of S.aureus isolates generated with primer GEN1-50-01 (5’GTGCAATGAG-3’) resulting in several polymorphic bands and a dendrogram was created. Conclusions: S. aureus is an important cause of HCAI, Health care workers represent an important source of HCAI, RAPD –PCR technique was an easy and rapid one to perform with a good discriminatory power in typing (fingerprinting) of Staphylococcus aureus.

We evaluated the specificity of the primers OPF-01, P54, and 1253 to identify A. fumigatus, A. flavus, and A. niger, respectively, with the RAPD-PCR method. Eighty-two isolates belonging to the sections Fumigati, Flavi, and Nigri were used. The isolates were identified by phenotypic (macro- and micromorphology) and genotypic (partial sequences of the BenA gene) methods. The RAPD-PCR method was used to obtain polymorphic patterns with the primers OPF-01, P54, and 1253. The specificity of the polymorphic patterns of the isolates of each species was evaluated through the UPGMA clustering method and logistic regression model. All isolates of the genus Aspergillus were identified at the section level by macro- and micromorphology showing the typical morphology of the sections Fumigati, Flavi, and Nigri, and the species were identified by the construction of the phylogeny of the partial sequence of the BenA gene. The patterns’ polymorphic strains obtained with the primers OPF-01, P54, and 1253 for the isolates of A. fumigatus, A. flavus, and A niger, respectively, showed the same polymorphic pattern as the reference strains for each species. To verify the specificity of the primers, they were tested with other species from the sections Fumigati, Flavi and Nigri. The results support that the primers OPF-01, P54, and 1253 generate polymorphic patterns by RAPD-PCR species specific to A. fumigatus, A. flavus, and A. niger, respectively.

Feta cheese is the most recognized Greek Protected Designation of Origin (PDO) product in the world. The addition of selected autochthonous lactic acid bacteria (LAB) strains to cheese milk as adjunct cultures is gaining more attention, since they can impact the nutritional, technological and sensory properties of cheeses, as well as improve the safety of the product. The aim of this study was to produce Feta cheese with enhanced quality and safety, and distinctive organoleptic characteristics by applying autochthonous lactic acid bacteria (LAB) with multi-functional properties as adjunct cultures. Feta cheeses were produced with the commercial lactococcal starter culture and the addition of 9 LAB strains (Lactococcus lactis SMX2 and SMX16, Levilactobacillus brevis SRX20, Lacticaseibacillus paracasei SRX10, Lactiplantibacillus plantarum FRX20 and FB1, Leuconostoc mesenteroides FMX3, FMX11, and FRX4, isolated from artisanal Greek cheeses) in different combinations to produce 13 cheese trials (12 Feta trials with the adjunct LAB isolates and the control trial). In addition, Feta cheese manufactured with FMX3 and SMX2 and control Feta cheese were artificially inoculated (4 log CFU/g) with Listeria monocytogenes (a cocktail of 4 acid or non-acid adapted strains). Cheese samples were monitored by microbiological and physicochemical analyses during ripening, and microbiological, physicochemical, molecular and sensory analyses during storage at 4°C. The results showed that after manufacture, the LAB population was ca. 9.0 log CFU/g at all samples, whereas during storage, their population declined to 6.5-7.0 log CFU/g. In the Listeria inoculated samples, Listeria was absent after 60 days (end of ripening) and after 90 days in the adjunct culture, and in the control trials, respectively. Moreover, the addition of selected strains, especially Lcb. paracasei SRX10, led to cheeses with desirable and distinctive organoleptic characteristics. Furthermore, randomly amplified polymorphic PCR (RAPD-PCR) molecular analysis confirmed that the multi-functional LAB strains were viable by the end of storage. Overall, the results of this study are promising for the use of autochthonous strains in various combinations with the commercial starter culture to satisfy industry requirements and consumer demands for traditional and high added value fermented products. [ABSTRACT FROM AUTHOR]

Simple Summary: Evaluating age-based differences in the resident lactic acid bacteria in the gut of healthy animals is important for potential probiotic species identification and development. This gives an understanding of unique bacterial populations that are well-adapted for survival in each age group. In goats, the resident gut lactic acid bacteria diversity in different age groups has not been fully described. The objective of the study was to evaluate the abundance and identity of the lactic acid bacteria populations in different age groups of goats from birth until ten months age. We detected higher counts of lactic acid bacteria in young goat kids before weaning than goats after weaning. Additionally, we detected different lactic acid bacteria populations in the different age groups evaluated. The findings give insight on potential lactic acid bacteria species that could be targeted for the development of probiotics for different ages of goats. These results may also broadly imply that when selecting potential bacteria for probiotic evaluation in other host animals, it is important to consider the target age to ensure success of establishment in the gut. Understanding gut lactic acid bacteria (LAB) in healthy hosts is an important first step in selecting potential probiotic species. To understand the dynamics of LAB in healthy goats, a cohort of thirty-seven healthy new-born goat kids was studied over a ten-month period. Total LAB was quantified using SYBR green qPCR. Seven hundred LAB isolates were characterized using microscopy, M13 RAPD genotyping and 16S rDNA sequencing. The highest and lowest LAB counts were detected at one week and ten months of age, respectively. Diverse LAB species were detected, whose identity and prevalence varied with age. The main isolates belonged to Limosilactobacillus reuteri, Limosilactibacillus fermentum, Lactobacillus johnsonni, Ligilactobacillus murinus, Ligilactobacillus salivarius, Limosilactobacillus mucosae, Lactiplantibacillus plantarum, Ligilactobacillus agilis, Lactobacillus acidophilus/amyolovolus, Pediococcus spp. and Enterococcus spp. Uniquely, L. reuteri and Pediococcus spp. were most common in pre- and peri-weaned goats, while Lactobacillus mucosae and Enterococcus spp. were predominant in goats one month and older. Based on RAPD genotyping, L. reuteri had the highest genotypic diversity, with age being a factor on the genotypes detected. This data may be relevant in the selection of age-specific probiotics for goats. The findings may also have broader implications by highlighting age as a factor for consideration in probiotic bacteria selection in other animal hosts. [ABSTRACT FROM AUTHOR]

Background: Healthcare-associated infections appear in a patient receiving medical attention at a hospital or other healthcare facility and develops infections that weren't present when they were admitted. One of the most important organisms causing infections acquired in hospitals is Staphylococcus aureus (S. aureus). For many bacterial species, Random Amplified Polymorphic DNA (RAPD) is a low-cost but effective typing technique. Objectives: To determine the genotypic polymorphism, the level of genetic relatedness, and the antimicrobial sensitivity and resistance among various S. aureus isolates. Methods: The following tests were performed on 50 samples that were obtained using strict aseptic precautions: sample culture, isolate identification using colony morphology, Gram stained film, biochemical responses, and antibiotic susceptibility testing. After RAPD-PCR, Syngene gene tool software was used to analyse the fingerprint pattern. Results: The most prevalent isolated organism was S. aureus. as it represented (70%) of isolated pathogens. Vancomycin and linezolid were effective against S. aureus isolates. The isolates of S. aureus were more common in nasal swab (42.9%) followed by wound (28.5%) then urine &endotracheal aspirate (14.3%). Patterns of RAPD-PCR of S.aureus isolates generated with primer GEN1-50-01 (5'GTGCAATGAG-3') resulting in several polymorphic bands and a dendrogram was created. Conclusions: S. aureus is an important cause of HCAI, Health care workers represent an important source of HCAI, RAPD -PCR technique was an easy and rapid one to perform with a good discriminatory power in typing (fingerprinting) of Staphylococcus aureus. [ABSTRACT FROM AUTHOR]

In this study, molecular typing using Randomly Amplified Polymorphic DNA (RAPD-PCR) was conducted on 16 original isolates of Metarhizium acridum obtained from locusts (Schistocerca piceifrons ssp. piceifrons.) in Mexico (MX). The analysis included reference strains of the genus Metarhizium sourced from various geographical regions. The isolates were identified by phenotypic (macro and micromorphology) and genotypic methods (RAPD-PCR and Amplified Fragment Length Polymorphisms (AFLP), through a multidimensional analysis of principal coordinates (PCoA) and a minimum spanning network (MST). Subsequently, Sequences-Characterized Amplified Region (SCAR) markers were developed for the molecular detection of M. acridum, these markers were chosen from polymorphic patterns obtained with 14 primers via RAPD-PCR. Phenotypic and genotypic characterization identified the MX isolates as M. acridum. Of all the polymorphic patterns obtained, only OPA04 and OPA05 were chosen, which presented species-specific bands for M. acridum, and further utilized to create SCAR markers through cloning and sequencing of the specific bands. The specificity of these two markers was confirmed via Southern hybridization. The SCAR markers (Ma-160OPA-05 and Ma-151OPA-04) exhibit remarkable sensitivity, detecting down to less than 0.1 ng, as well as high specificity, as evidenced by their inability to cross-amplify or generate amplification with DNAs from other strains of Metarhizium (as Metarhizium anisopliae) or different genera of entomopathogenic fungi (Cordyceps fumosorosea and Akanthomyces lecanii). These SCAR markers yield readily detectable results, showcasing high reproducibility. They serve as a valuable tool, especially in field applications.

Ornithobacterium rhinotracheale (ORT) is a newly emerging respiratory bacterial pathogen that causes significant economic losses to the poultry industry. Previous studies in Iran have shown high molecular and genetic similarity among ORT isolates by SDS-PAGE, ERIC-PCR and 16SrRNA gene sequencing during 1999-2009. The aim of this study was the genotyping of ORT isolates recovered from commercial chickens by using RAPD-PCR with OPG11 primer and multilocus sequence typing (MLST). In total, 30 ORT isolates recovered from commercial chickens of Iran during 2000-2017 and confirmed by bacteriological, biochemical and PCR tests were used in this study. All 30 ORT isolates were subjected to RAPD-PCR with OPG11 primer. For MLST, 5 isolates were selected based on their RAPD patterns. Seven primer pairs were synthesized for amplification and sequenceing of seven housekeeping genes of adk, aroE, fumC, gdhA, mdh, pgi and pmi in MLST assay. In RAPD-PCR with OPG11 primer, 9 different genotypes were found. The DNA sequences of the distinct alleles of these seven loci of 5 ORT strains were compared with other alleles deposited in GenBank. Four out of 5 strains belonged to sequence type 9 (ST9) and one strain was found to be a new ST. Based on the results of the present study, a new sequence type among ORT isolates of Iran was found that has not been previously reported from elsewhere in the world. Further studies on more ORT isolates may help in identification of different sequence types and dominat ST in the country. [ABSTRACT FROM AUTHOR]

The family Cactaceae comprises many genera, has taxonomic controversies at the species level, and yet has not been studied at the genetic level in Iraq. The presented study focused on species genotyping based on the RAPD-PCR method. The classification of six species, i.e., Hylocereus undatus, Aloe vera, Opuntia ficus-indica, Espostoa guentheri, Echinocactus grusonii, and Mammillaria elongata, ensued based on phenotypic characters to determine their precise taxonomic names and evaluate kinship by constructing the phylogeny tree RAPD-PCR. The short oligo primers showed the highest polymorphic bands (100%), with no monomorphic and basal bands correlated among these species. This highly polymorphic relationship indicated that each species has a superior identity and unique evolutionary trend. The oligo primers were considered productive by showing highly distinct and sharp bands, while others showed faint bands. This research confirmed the efficacy of RAPD primers in measuring polymorphism, comparing genotypes, and identifying Cacti species using specialized RAPD markers. [ABSTRACT FROM AUTHOR]

The objective of this study was to identify morphological and molecular comparison of three marine Chaetoceros species using microscopic observations, sequence analysis of 18S rDNA, random amplified polymorphic DNA (RAPD-PCR) barcoding and nuclear magnetic resonance (NMR) spectroscopy. Chaetoceros were obtained from three different algae laboratories: Center of Excellence for Marine Biotechnology (CEMB), Chanthaburi Coastal Fisheries Research and Development (CHAN) and Institute of Marine Science, Burapha University (BIM). Genomic DNA for the RAPD-PCR analysis was extracted using the phenol–chloroform method, followed by 18S rDNA amplification. The blast results of 18S rDNA sequence confirmed the significantly matched to C. gracilis for Chaetoceros BIM and CHAN and C. muelleri for Chaetoceros CEMB(e-value = 0.0, identity = 99%). The RAPD-PCR results revealed differences in the three Chaetoceros isolates with polymorphisms between 30.43% and 60.00%, and Chaetoceros CEMB showed high polymorphic bands. Scanning electron microscopy revealed that Chaetoceros CEMB were larger and had larger setae compared to the other isolates (P < 0.05). The results of the NMR characterization of metabolites were consistent with the results of the sequence and morphological analyses. The concentrations of several metabolites, including chlorophyll c1, chlorophyll a, Myo-inositol, fucoxanthin, astaxanthin, lutein and zeaxanthin, were lower in Chaetoceros CEMB than in Chaetoceros BIM and CHAN. However, high concentrations of fatty acids, such as oleic acid, linoleic acid, α-linolenic acid and arachidic acid, were observed in all isolates. Generally, the results of this study will aid future studies examining the diversity of Chaetoceros in various cultural environments. [ABSTRACT FROM AUTHOR]

Abstract Background The Prototheca algae have recently emerged as an important cause of bovine mastitis globally. Isolates from bovine mastitis in several countries were nearly all identified as P. bovis, suggesting that it was the main causative agent of bovine protothecal mastitis. The aim of the present study was to evaluate the presence and isolation of Prototheca spp. in dairy farms, detect the genetic diversity among strains, determine the capacity of producing biofilm and their resistance to antifungal and antimicrobial drugs. Results A total of 48 Prototheca isolates from four different farms were randomly selected to be investigated. Multiplex PCR showed all isolated colonies were Prototheca bovis. Performing RAPD-PCR by using OPA-4 primer, it was revealed that there was a clear amplification pattern. Different levels of biofilm production were observed among strains. Among 48 isolates, only 4 of them (8.33%) showed strong biofilm production. By using E-test strips, amphotericin B was able to inhibit the growth of all the strains tested. Disc diffusion method used for antimicrobial sensitivity test showed that the highest activity was demonstrated by gentamicin and colistin with 95.83% (46/48) and 89.58% (43/48) of sensitive strains, respectively. Conclusions The present study showed that RAPD-PCR was a rapid tool for discriminating P. bovis strains. Also, gentamicin and colistin can be considered as potential antimicrobial drugs which can prevent the growth of the mentioned strains in vitro, although there is no effective clinical treatment yet. Further studies are needed in order to detect an effective clinical therapy considering biofilm production by Prototheca spp. and their probable role in Prototheca pathogenicity.

Antimicrobial therapy is frequently associated with the emergence of resistant bacteria with a high rate of morbidity and mortality worldwide. The present study was aimed at investigating the impact of a neodymium-doped yttrium aluminum (Nd:YAG) laser, and a static magnetic field (SMF) on cellular growth and DNA alteration in some clinical bacterial isolates. Samples from cutaneous wounds were collected by sterile cotton swabs from three elderly women admitted to Tikrit Teaching Hospital, Tikrit City, Iraq. Isolation and identification of Streptococcus agalactiae, Staphylococcus aureus, and Pseudomonas aeruginosa were carried out using cultural characteristics, microscopy, and biochemical tests. Three broth cultures were prepared for each of the test isolates. The first broth culture served as untreated control, the second was exposed to an Nd:YAG laser and the third was exposed to SMF. Colony counting was done on all the samples. DNA was extracted from the test bacteria and used to perform the RAPD-PCR assay. In contrast to the untreated control, the results showed that Nd:YAG laser radiation was more effective than SMF at inhibiting the cellular growth of the test isolates. Also, the radiation caused DNA alteration, which was established by decreased microbial growth, as well as the development of new bands and the loss of original bands. According to the findings of this study, the Nd:YAG laser is a promising technique for influencing the healing of infected cutaneous wounds. RAPD-PCR is also a useful biomarker assay for assessing the biological impact of laser radiation and SMF on bacteria.

Recently, methods based on the analysis of arbitrarily amplified target sites of genome microorganisms have been extensively applied in microbiological studies, and especially in epidemiological studies. The range of their application is limited by problems with discrimination and reproducibility resulting from a lack of standardized and reliable methods of optimization. The aim of this study was to obtain optimal parameters of the Random Amplified Polymorphic DNA (RAPD) reaction by using an orthogonal array as per the Taguchi and Wu protocol, modified by Cobb and Clark for Candida parapsilosis isolates. High Simpson's index values and low Dice coefficients obtained in this study indicated a high level of interspecies DNA polymorphism between C. parapsilosis strains, and the optimized RAPD method proved useful in the microbiological and epidemiological study. [ABSTRACT FROM AUTHOR]

Understanding the actual distribution of different Legionella species in water networks would help prevent outbreaks. Culture investigations followed by serological agglutination tests, with poly/monovalent antisera, still represent the gold standard for isolation and identification of Legionella strains. However, also MALDI-TOF and mip-gene sequencing are currently used. This study was conducted to genetically correlate strains of Legionella non pneumophila (L-np) isolated during environmental surveillance comparing different molecular techniques. Overall, 346 water samples were collected from the water system of four pavilions located in a hospital of the Apulia Region of Italy. Strains isolated from the samples were then identified by serological tests, MALDI-TOF, and mip-gene sequencing. Overall, 24.9% of water samples were positive for Legionella, among which the majority were Legionella pneumophila (Lpn) 1 (52.3%), followed by Lpn2-15 (20.9%), L-np (17.4%), Lpn1 + Lpn2-15 (7.1%), and L-np + Lpn1 (2.3%). Initially, L-np strains were identified as L. bozemanii by monovalent antiserum, while MALDI-TOF and mip-gene sequencing assigned them to L. anisa. More cold water than hot water samples were contaminated by L. anisa (p < 0.001). PFGE, RAPD, Rep-PCR, and SAU-PCR were performed to correlate L. anisa strains. Eleven out of 14 strains identified in all four pavilions showed 100% of similarity upon PFGE analysis. RAPD, Rep-PCR, and SAU-PCR showed greater discriminative power than PFGE. [ABSTRACT FROM AUTHOR]

The confused flour beetle, Tribolium confusum, is a common insect that can be found in stores and homes and attacks a variety of commodities and stored foods. In the past, synthetic chemical insecticides were used to control them, which caused many negative health and environmental effects. Acetamiprid is a neonicotinoid insecticide with a novel mechanism of action, low toxicity for mammals, and a low environmental impact. The effectiveness of that insecticide against T. confusum was tested in this study. The toxicological bioassay was performed by contact application with different concentrations of acetamiprid. Our results indicated that the increased concentration and exposure periods significantly enhanced the mortality rate. The mortalities reached 20.6±1.12% and 92.6±1.86% at 1 ppm and 8 ppm concentrations of acetamiprid, respectively, after 14 days of exposure. The effects of acetamiprid on the fecundity and sterility of T. confusum have been assessed. Laboratory studies indicated that acetamiprid significantly reduced egg laying and hatching in comparison with the control. All acetamiprid concentrations used resulted in sterility in the treated adults. At the highest concentration (8 ppm), the sterility was 66.28%. The effects of acetamiprid on the activity of the acetylcholinesterase (AChE) enzyme were also investigated. It was observed that AChE from T. confusum was almost completely inhibited at the highest concentration. On the contrary, the level of the detoxifying enzyme, glutathione S-transferase, was increased in the treated samples. Acetamiprid's mutagenic effect on T. confusum was investigated by applying the median-lethal concentration (LC50), investigating DNA alteration for polymorphic numbers of genetic bands by using RAPD-PCR primers, and comparing it with the control. Results strongly suggest that acetamiprid causes mutagenic effects on T. confusum. The results recommended that acetamiprid can be used as a useful tool in pest management programs for T. confusum considering its insecticidal effects, which are evident at low applied concentrations and short exposure times. [ABSTRACT FROM AUTHOR]

Cerium oxide engineered nanoparticles (nCeO2) are widely used in various applications and are, also, increasingly being detected in different environmental matrixes. However, their impacts on the aquatic environment remain poorly quantified. Hence, there is a need to investigate their effects on non-target aquatic organisms. Here, we evaluated the cytotoxic and genotoxic effects of <25 nm uncoated-nCeO2 on algae Pseudokirchneriella subcapitata. Apical (growth and chlorophyll a (Chl a) content) and genotoxic effects were investigated at 62.5–1000 µg/L after 72 and 168 h. Results demonstrated that nCeO2 induced significant growth inhibition after 72 h and promotion post 96–168 h. Conversely, nCeO2 induced enhanced Chl a content post 72 h, but no significant changes were observed between nCeO2–exposed and control samples after 168 h. Hence, the results indicate P. subcapitata photosynthetic system recovery ability to nCeO2 effects under chronic-exposure conditions. RAPD-PCR profiles showed the appearance and/or disappearance of normal bands relative to controls; indicative of DNA damage and/or DNA mutation. Unlike cell recovery observed post 96 h, DNA damage persisted over 168 h. Thus, sub-lethal nCeO2-induced toxicological effects may pose a more serious threat to algae than at present anticipated. [ABSTRACT FROM AUTHOR]

The aim of this study was ‎the‎ discrimination of Salmonella‎‎ isolated from chicken and their feed ‎and drinking water for the epidemiological control of salmonellosis. Totally, 289 samples, ‎including 217 chicken cloaca swabs, 46 water, and 26 feed samples were collected from five ‎different farms in Karbala governorate, Iraq. Conventional bacteriology tests, API 20E, Vitek 2, ‎and serology were used for bacterial identification. Random amplified polymorphic ‎DNA (RAPD)-polymerase chain reaction (PCR) was applied to analyze the genetic relationships ‎among Salmonella‎‎ isolates. The isolation rate of Salmonella‎‎ spp. was 21.1% (61/289). While the ‎water samples constituted the highest rate (30.4%), a rate of 21.7% was reported for the cloaca ‎swabs, with no isolate at all from chicken feed. Vitek 2 was able to identify some isolates to the ‎serotype level, such as S. Enteritidis, S. Paratyphi B, and S. Paratyphi C. However, the isolates ‎were diagnosed as S. enterica by API 20E, and as S. enterica subsp. arizonae through serology. ‎Analyzing the samples by the RAPD-PCR assay showed the presence of genetically different ‎Salmonella‎‎ spp. Dendrograms created by the GelJ software successfully delineated the genetic ‎relationships. Therefore, RAPD-PCR can be used as a surrogate tool for the fast, reliable, and ‎accurate detection of Salmonella‎‎ in epidemiological surveys when compared with other ‎biochemical-based identification methods.

Objetivos: Conocer la diversidad genética de Aedes aegypti en el corredor vial transfronterizo Central- Alto Paraná de Paraguay, con registros de casos de dengue. Materiales y métodos: Se seleccionaron veinte hembras adultas de la eclosión de huevos de Ae. aegypti procedentes de casas geolocalizadas en los departamentos de Alto Paraná, Caaguazú, Cordillera y Central, entre el 2018 y 2019. Se extrajo ADN del tejido de las hembras para amplificación aleatoria de sus patrones polimórficos mediante amplificación aleatoria del ADN polimórfico por PCR (RAPD-PCR), usando cebadores H3 y B03 a fin de conocer parámetros genéticos de diversidad poblacional. Las relaciones entre las poblaciones de mosquitos según la localidad fueron visualizadas mediante análisis no apareado de la media aritmética. Las áreas idóneas de distribución geográfica real y potencial de estas poblaciones de Ae. aegypti fueron analizadas mediante DIVA-GIS 7.3.0 y MAXENT. Resultados: Se identificaron 40 loci mediante perfiles RAPD-PCR, con diferenciación génica moderada (Gst = 0,12). El corredor transfronterizo presentó condiciones bioclimáticas para la presencia de poblaciones variantes de Ae. aegypti, siendo determinantes en la distribución la precipitación del trimestre más cálido y la temperatura media del trimestre más seco. Conclusiones: Se evidencia que existe diversidad genética moderada en las poblaciones de Ae. aegypti procedentes de zonas con registros de casos de dengue ubicadas en el corredor vial transfronterizo que une los departamentos Central y Alto Paraná de Paraguay. El estudio de variabilidad genética de Ae. aegypti es de gran utilidad para la vigilancia entomoepidemiológica y evaluación de posibles eventos de resistencia al control químico.

Copyright of Iraqi Journal of Veterinary Medicine is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: The Prototheca algae have recently emerged as an important cause of bovine mastitis globally. Isolates from bovine mastitis in several countries were nearly all identified as P. bovis, suggesting that it was the main causative agent of bovine protothecal mastitis. The aim of the present study was to evaluate the presence and isolation of Prototheca spp. in dairy farms, detect the genetic diversity among strains, determine the capacity of producing biofilm and their resistance to antifungal and antimicrobial drugs. Results: A total of 48 Prototheca isolates from four different farms were randomly selected to be investigated. Multiplex PCR showed all isolated colonies were Prototheca bovis. Performing RAPD-PCR by using OPA-4 primer, it was revealed that there was a clear amplification pattern. Different levels of biofilm production were observed among strains. Among 48 isolates, only 4 of them (8.33%) showed strong biofilm production. By using E-test strips, amphotericin B was able to inhibit the growth of all the strains tested. Disc diffusion method used for antimicrobial sensitivity test showed that the highest activity was demonstrated by gentamicin and colistin with 95.83% (46/48) and 89.58% (43/48) of sensitive strains, respectively. Conclusions: The present study showed that RAPD-PCR was a rapid tool for discriminating P. bovis strains. Also, gentamicin and colistin can be considered as potential antimicrobial drugs which can prevent the growth of the mentioned strains in vitro, although there is no effective clinical treatment yet. Further studies are needed in order to detect an effective clinical therapy considering biofilm production by Prototheca spp. and their probable role in Prototheca pathogenicity. [ABSTRACT FROM AUTHOR]

Agarose gel electrophoresis is a well-known tool to detect DNA fragments amplified in polymerase chain reaction (PCR). Its usefulness has also been confirmed for epidemiological studies based on restriction fragments length polymorphism (RFLP), usually performed using pulsed-field gel electrophoresis (PFGE). Little is known on the effectiveness for alert-pathogen epidemiological studies of another less time-consuming and costly technique called randomly amplified polymorphic DNA-PCR (RAPD-PCR). Meanwhile, its usefulness is believed to be comparable to RFLP-PFGE. Therefore, the aim of the study was to establish and optimize the conditions of agarose gel electrophoresis following RAPD-PCR for 19 Enterococcus faecium strains derived from epidemic outbreaks at intensive care units. An application of different PCR primers, primer combinations, and, in particular, agarose gel concentrations and electrophoresis conditions revealed the usefulness of this relatively fast and inexpensive method based on RAPD-PCR for epidemiological studies without a compulsion to use the specialized equipment necessary for RFLP-PFGE. [ABSTRACT FROM AUTHOR]

A wide range of options for studying Candida species are available through genetic methods. Twins, particularly monozygotic ones and their families may be fitting subjects for studying those microorganisms. The question is: How specific can yeast flora be in an individual? The study aimed to analyze the strain relatedness among commensal yeasts isolated from various parts of the bodies of healthy people and to compare correlations between the genotypes of the isolates. Yeasts were isolated from 63 twins and their family members (n = 25) from the oral cavity, anus, interdigital space and navel. After species identification, Candida albicans (n = 139), C. parapsilosis (n = 39), C. guilliermondii (n = 25), C. dubliniensis (n = 11) and C. krusei (n = 9) isolates were analyzed using the random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) optimization method. The similarities between the strains were calculated based on the Dice (Sab) coefficient and are displayed graphically as dendrograms. Using cluster analysis, the following relatedness was distinguished: 13 genotypes and three unique (Un) patterns among C. albicans; 10 genotypes and four Un patterns among C. parapsilosis; three genotypes and one Un pattern among C. guilliermondii and C. dubliniensis; and three genotypes among C. krusei isolates. The presence of identical, similar or both genotypes among the strains isolated from family members shows the transmission of yeasts between ontocenoses in the same person and between individuals. The similarity between the genotypes of C. albicans, C. guilliermondii, C. dubliniensis and C. krusei was more remarkable than between the genotypes of C. parapsilosis in the strains isolated from ontocenoses of the same individual and their family members. The degrees of genetic similarity between Candida spp. strains isolated from monozygotic twins and those obtained from their relatives did not differ. [ABSTRACT FROM AUTHOR]

Copyright of Baghdad Science Journal is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Purpose: The aim of this study as to compare microscopy and culture results of samples, determine drug resistance rates of the isolates, evaluate epidemiological relationship between the strains with ERIC-PCR, RAPD-PCR, OUT-PCR based on in house PCR technique. Materials and Methods: Direct microscopy and culture results of 2010 samples were analyzed. Drug sensitivity results were obtained from TULSA. The typing of isolates based on in house PCR was carried out in the microbiology laboratory of a faculty of medicine in a state university. Results: Of positive samples, 2.68% had Acid-resistant bacilli (ARB) positive + culture positivity, 2.93% had ARB positive and culture negative in smear, and 1.34% had ARB negative and culture positivity in smear. Resistance to primary antituberculous (anti-TB) drugs wasn’t observed in 33 culture positive isolates, whereas resistance to one or more primary anti-TB drugs was observed in 9.09%. Single drug resistance was 3.03%, resistance to Isoniazid (INH) and INH critical drugs was 6.06%. Isolates were divided 3 groups by ERIC-PCR, 5 groups for OUT-PCR and 6 by RAPD-PCR. Conclusion: Positivity rates were low due to low rate of studied samples, negative samples taken during treatment process. Molecular techniques like ERIC-PCR, RAPD-PCR and OUT-PCR are easy, fast and inexpensive methods for the epidemiological typing of Mycobacterium tuberculosis (MTB) in evaluating distinctions, similarities between origins.

Bacillus bacteria are advantageous antagonistic organisms that can be used as bio-control agents. This study is aimed at screening the antagonistic activity of different strains of isolated Bacillus bacteria and molecular identification of the superior chitinase producer strain against dermatophytes fungi. Soil samples were collected from different places of Kotoor city, Gharbia Governorate, Egypt and Al Madina Al Munawwarah, Kingdom of Saudi Arabia. A collection of Bacillus isolated from soil was tested in vitro against the dermatophytes: Microsporum sp. and Trichophyton sp. The bacterial strains Kh-B1 and Kh-B2 showed the highest antagonistic activity against dermatophytes pathogenic fungi. The highest amount of chitinase productivity (13.6 units/ml) was obtained from the original Bacillus strain (Kh-B1) at 3 days of incubation. BLAST analysis of the amplified 16S ribosomal RNA gene sequence identified the Bacillus strain (Kh-B1) as Paenibacillus macerans. Upon the mutation induction by UV light, the highest chitinase-producing mutant was Kh-UVB-4 as it showed 305.88 percent production higher than the wild-type strain. While, upon the mutation induction by EMS, the highest amount of chitinase produced was 54.8 units/ml by mutant Kh-ESB-20, and it has produced 402.94% more than the original untreated strain. The application of RAPD-PCR protocol using three 15-mer random primers was used to determine the genetic effects of mutagenic treatments on the wild type strain (Kh-B1) as well as to demonstrate the genetic variability between the five most chitinase producing mutants and the wild type (Paenibacillus macerans).

Understanding gut lactic acid bacteria (LAB) in healthy hosts is an important first step in selecting potential probiotic species. To understand the dynamics of LAB in healthy goats, a cohort of thirty-seven healthy new-born goat kids was studied over a ten-month period. Total LAB was quantified using SYBR green qPCR. Seven hundred LAB isolates were characterized using microscopy, M13 RAPD genotyping and 16S rDNA sequencing. The highest and lowest LAB counts were detected at one week and ten months of age, respectively. Diverse LAB species were detected, whose identity and prevalence varied with age. The main isolates belonged to Limosilactobacillus reuteri, Limosilactibacillus fermentum, Lactobacillus johnsonni, Ligilactobacillus murinus, Ligilactobacillus salivarius, Limosilactobacillus mucosae, Lactiplantibacillus plantarum, Ligilactobacillus agilis, Lactobacillus acidophilus/amyolovolus, Pediococcus spp. and Enterococcus spp. Uniquely, L. reuteri and Pediococcus spp. were most common in pre- and peri-weaned goats, while Lactobacillus mucosae and Enterococcus spp. were predominant in goats one month and older. Based on RAPD genotyping, L. reuteri had the highest genotypic diversity, with age being a factor on the genotypes detected. This data may be relevant in the selection of age-specific probiotics for goats. The findings may also have broader implications by highlighting age as a factor for consideration in probiotic bacteria selection in other animal hosts.

This study aimed at isolating uropathogenic Escherichia coli from urinary tract infections (UTIs) of human and cattle to examine the molecular diversity and phylogenetic relationship of the isolates. A total of 100 urine samples were collected from UTIs of human and cattle. The isolates identification was done using routine diagnostic methods and confirmed by Vitek2. Antimicrobial susceptibility was tested against 10 antimicrobials. Random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) was applied to identify the genetic diversity among E. coli isolates from human and animal origin by using five different octamer primers. The gelJ software for the phylogenetic analysis created Dendrograms. Out of 50 human urine samples, E. coli was isolated from 12 (24%) samples, and was positive in 5 out of 50 (10%) of cattle urine samples. Concerning the antimicrobial susceptibility test, both human and animal isolates revealed rather approximate results when tested mainly against Imipenem, Cefotaxime, and Ciprofloxacin. These antimicrobial data might indicate presence of a degree of similarity between the human and animal isolates. Using RAPD-PCR, three of the primers produced polymorphic bands; therefore, they were used for further analysis of the results. Either of P1, P3 or P4 primers showed presence of similarity between human and cow isolates. To conclude,RAPD-PCR and gelJ software might be of attractive use to identify and analyze the occurrence of genetic relationships, as this could assist in controlling the routes and sources of infection transmission between human and animal in order to prevent zoonotic infections.