Introduction
Raw milk is a nutrient‐rich food, but it may harbour harmful bacteria, such as enterotoxigenic Staphylococcus aureus (S. aureus), which can cause staphylococcal food poisoning. ...Antibiotic resistance of S. aureus in raw milk can increase the risk of such infections, particularly among susceptible individuals.
Objective
This study aimed to investigate the prevalence of enterotoxin genes a, d, g, i and j and the antibiotic resistance of S. aureus isolated from raw milk samples.
Methods
During a 6‐month sampling period, 60 raw milk specimens were obtained from diverse locations in Yazd province, Iran. Antibiogram profiling was conducted via the disc diffusion method. In addition, staphylococcal enterotoxin (SE) genes a, d, g, i, and j were detected through real‐time PCR analysis.
Results
Bacteriological assays confirmed the presence of S. aureus in 11 samples (18.3%). All isolates demonstrated 100% resistance to penicillin G but exhibited sensitivity to vancomycin, while resistance to other antibiotics ranged from 36.4% to 45.5%. The prevalence of enterotoxin genes in these strains showed variable distribution, with sea being the predominant SE (45.5%), followed by sed (36.4%), seg (18.2), sej and sei (9.1% each).
Conclusions
This study discovered the presence of multiple enterotoxins in S. aureus strains obtained from raw milk samples. These strains also demonstrated resistance to a variety of antibiotics. Since enterotoxigenic S. aureus is known to cause human food poisoning, monitoring food hygiene practices, especially during raw milk production, is critical.
In this study culture, biochemical tests, real‐time PCR, and the Kirby‐Bauer disk diffusion method were used to identify and characterise the isolates. Enterotoxin genes and antibiotic resistance were detected in 11 (18.3%) S. aureus strains from raw milk samples. The strains had the sea, sed, seg, sei and sej genes, with the sea being the most frequent. All strains resisted penicillin G but were susceptible to vancomycin. Resistance to other antibiotics ranged from 36.4% to 45.5%.
High-resolution melting (HRM) is a reliable and sensitive scanning method to detect variation in DNA sequences. We used this method to better understand the epidemiology and transmission of ...Echinococcus granulosus. We tested the use of HRM to discriminate the genotypes of E. granulosus and E. canadensis. One hundred forty-one hydatid cysts were collected from slaughtered animals in different parts of Isfahan-Iran in 2013. After DNA extraction, the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was amplified using PCR coupled with the HRM curve. The result of HRM analysis using partial the sequences of cox1 gene revealed that 93, 35, and 2 isolates were identified as G1, G3, and G6 genotypes, respectively. A single nucleotide polymorphism (SNP) was found in locus 9867 of the cox1 gene. This is a critical locus for the differentiation between the G6 and G7 genotypes. In the phylogenic tree, the sample with a SNP was located between the G6 and G7 genotypes, which suggest that this isolate has a G6/G7 genotype. The HRM analysis developed in the present study provides a powerful technique for molecular and epidemiological studies on echinococcosis in humans and animals.
Determination of the prevalence of intestinal protozoan infection is a fundamental step to set up an effective control program to improve the health status of society and to establish efficient ...strategies. Intestinal pathogen and even non-pathogen protozoa consider as major causes of disease in patients with gastrointestinal problems. The objective of this study is to determine the prevalence of intestinal protozoan infection in patients with ulcerative colitis (UC) in Isfahan, Iran.
The descriptive cross-sectional study carried out from 2013 to 2018 in Isfahan, Iran. One thousand nine hundred and sixty-five samples of feces from patients with UC collected and each sample examined using direct wet mounting with normal saline and iodine and sedimentation tests such as formol-ethyl acetate concentration and trichrome-staining methods.
From 655 patients, 185 (28.2%) infected with
followed by
(27.3%),
(14.4%),
(11.5%),
(4.7%),
(1.4%), and
(0.6%).
This study revealed a high prevalence of infection with at least one or six non-pathogenic and pathogenic intestinal protozoa in UC patients in the Isfahan region. Intestinal protozoa are a challenging public health problem wherever health care is limited in the area. The emergence of UC in the world results in the need to study etiologic factors. In order to obtain further information about the etiology of disease, we investigated the prevalence of intestinal protozoan infection in patients with UC in Isfahan, Iran.
Toxoplasmosis is a zoonotic parasitic disease of global importance. It is widespread and endemic practically all over the world, with infection prevalence varying by geographic location. The parasite ...and the disease are neglected, which is illustrated by the lack of baseline information on the prevalence from many regions. Developed, peaceful regions are in better situation to address toxoplasmosis, while the neglected status is particularly pronounced in regions with other challenges. Due to the lack of baseline data, these regions are rarely mentioned in discussions about the neglected status of the disease. A dramatic manifestation of toxoplasmosis, congenital toxoplasmosis, is the dreaded outcome of vertical transmission of the infection from the mother to the unborn child. For this reason, pregnant women are a key target group for primary prevention of T. gondii infections, and baseline data on the prevalence in pregnant women is important. In this cross-sectional seroepidemiological study, we estimated Toxoplasma gondii seroprevalence and evaluated possible risk factors for seropositivity in pregnant women in Kabul, Afghanistan. Altogether 207 of the 431 women included in the study tested positive for immunoglobulin G antibodies against T. gondii, yielding an apparent seroprevalence estimate of 48.03% (95% CI 43.33-52.75). Based on the final multivariable model for T. gondii seropositivity, non-concrete floor in the house and well or river as water source were identified as risk factors for seropositivity, while residence in rural area was a protective factor. The majority of the participants (72.9%) reported that they did not know about T. gondii. Our study is the first to report an estimate of T. gondii seroprevalence in pregnant women in Afghanistan. The high seroprevalence indicates substantial infection pressure, and the results of the risk factor analysis suggest that the environmental route, infection from oocysts, might be the most relevant to address to prevent the infections in the region. Our results contribute to the global discussion on neglected status of toxoplasmosis.
The lactobacilli are abundant in honey, helping protect against pathogens and providing antimicrobial properties. This study aimed to isolate lactobacillus species from different honey regions and ...evaluate their potential probiotic properties.
Eighty-eight samples were collected from different regions, including the northern, central, and southern areas, and obtained through retail stores. All samples were independently examined for the presence of
using both culture and real-time PCR methods. Probiotic tests were performed on the isolated
strains, including hemolytic activity, bile, acid, and pepsin resistance. Additionally, the antibiotic resistance of the obtained strains was investigated using seven different antibiotics.
Thirteen
isolates were obtained from 7 (8.0%) honey samples. Of these, eight isolates were identified as
(61.54%), four isolates as
(30.77%), and one isolate as
(7.69%). All strains were devoid of hemolytic activity, and three isolates (23.07%) were found to be resistant to acid, while 2 (15.38%) showed resistance to bile and pepsin. All isolates were resistant to vancomycin (100%). Additionally, only one strain exhibited resistance to all tested antibiotics. Furthermore, the present study demonstrates a significant association (p-value<0.05) between the presence of
in various regions of Iran.
Various factors, such as climatic conditions and geographical location, can influence honey's composition and microbial diversity. Identifying and isolating potential probiotic species in honey could significantly expand their use in the food and pharmaceutical industries, offering numerous health benefits and potential therapeutic applications.
Among the human parasitic diseases, malaria is the main cause of morbidity and mortality. To prevent the high mortality and tracking malaria elimination efforts, a prompt and sensitive diagnosis is ...essential. This study aimed to compare High-Resolution Melting (HRM) and microscopic methods to diagnose
and
.
Eighty-one blood samples were collected from patients with clinical symptoms who were suspect to malaria in Chabahar district, southeastern Iran and also, from those who were referred to Malaria National Laboratory in the Tehran University of Medical Sciences, Tehran, Iran. Microscopic examination and HRM method were used to the diagnosis of
parasites simultaneously.
Microscopic results revealed 45 positive cases (12
and 33
) out of 81 collected samples while according to HRM analysis results 11 and 33 samples were identified as
and
, respectively. HRM analysis also revealed 1 mixed infection of
and
HRM analysis provides a promising mean for simultaneous detection and discrimination of the
spp. especially in mixed infection cases.
We aimed at genotyping and evaluating the predominance of
assemblages isolated from patients referred to medical laboratories in Khorramabad, Iran from Nov 2015 to Sep 2016. Hence, the development of ...a cost-effective HRM approach to determine genotypes of
based on the triosephosphate isomerase (tpi) gene was examined and the genotyping results with and without diarrhea was compared.
Seventy
positive fecal samples were collected. A microscopic confirmation for the presence of
spp. was performed, cysts of 70
spp. positive specimens were concentrated using sucrose flotation technique and sucrose solution PCR amplification was performed on 69 of 70 (98.5%) samples, and High Resolution Melting (HRM) analysis was performed using a software.
The results showed two distinct genotypes (assemblages A and B) of
but infections with mixture of both assemblages were not detected. The genotypes of
showed that the sub assemblage AI, BIII and BIV were present in a proportion of 68.1%, 20.3% and 11.6% respectively in samples. Assemblage AI was significantly (
<0.05) more frequently found in patients with diarrhea.
The sub-assemblage AI, BIII, and BIV are more zoonotic potential. According to the comparison of the results of this study with the results of previous studies in this area and around of it, as well as the way people live and keep pets. This pattern established in Khorramabad city. HRM can be an ideal technique to detect and genotyping of
in clinical samples.
Background: It is difficult to make an exact morphological distinction between Fasciola hepatica and Fasciola gigantica. We used High Resolution Melting analysis (HRM) method to differentiate the F. ...hepatica species from F. gigantica in order to differentiate them. Methods: Overall, 80 adult liver flukes were collected from infected slaughtered animals including cattle, sheep and goats from Lorestan Province, western Iran from Sep 2015 to Aug 2017. Genomic DNA was extracted using commercial DNA extraction kit. The multilocus sequences of mDNA including COX1, COX3 and ND6 were amplified employing real-time PCR & HRM analysis. Specific and universal primer pairs were designed for differentiation Fasciola spp. Results: Universal primers cannot be used to distinguish between these two species, but in the contrary, specific primer pairs of each species could differentiate them properly. Molecular identification using specific primer pairs were consistent. Conclusion: HRM is a simple, fast and reliable method for detecting and differentiating F. hepatica from F. gigantica and can be used for diagnostic and epidemiological purposes.
Leishmania (L.) infantum is the principal agent of visceral leishmaniasis (VL) in the Mediterranean and American regions. So far different molecular methods including high resolution melting (HRM) ...analysis have been developed for detecting and identifying L. infantum infection. HRM assay is an automted molecular method which detects and identifies different genus and species of infectious agents. This study aimed to diagnose and identify Leishmania infection caused by L. infantum species using real-time PCR coupled with HRM assay in the serum samples in comparison with anti-L. infantum antibodies obtained using direct agglutination test (DAT), in domestic and wild canines of northeastern Iran.
Serum samples of 15 foxes, 14 jackals, seven domestic dogs and three wolves were collected in some villages around Shirvan and Bojnourd districts from the northeast regions of Iran during 2014-15. Initially, all the collected serum samples were tested by DAT for the detection of anti-L. infantum antibodies. Afterwards, genomic DNA was extracted from the samples and tested by real-time PCR-HRM analysis targeting hsp70, ITS1 and gp63 genes. The level of agreement between DAT and HRM assay were analysed statistically.
Out of the 39 serum samples, eight showed anti-L. infantum antibodies at titre 1: 80 while only one of them showed anti-L. infantum antibodies at titre 1 : 160. All the nine seropositive samples showed positive results with HRM analysis. Additionally, three DAT negative serum samples were also found positive in the HRM technique. Altogether, 12 out of the 39 DNA samples showed positive results in HRM analysis. Among the three gene sequences used, gp63 was best for separation and identification of species.
HRM analysis targeting hsp70, ITS1 and gp63 genes can be used as a highly sensitive technique for the screening and early detection of L. infantum infection in the wild and domestic canines. It has higher accuracy than DAT and allows detection and discrimination of different Leishmania species responsible for the Leishmaniases.
Fasciolosis is a shared disease between humans and livestock caused by hepatic trematodes;
and
. Differentiate between the two species of this genus is essential. High-Resolution Melting (HRM) ...Analysis represents a new approach to this issue. This method can be performed right after termination of Real-Time PCR. This technique has not been used for identification of adult
and
genotypes. The aim of this study was to determine
species by using HRM in isolates taken from Iran, respectively.
Ninety-three
spp. samples were collected from infected slaughtered animals in different regions of Iran, including North West (Ardebil Province) and South East (Zahedan Province) during 2016. Genomic DNA from the samples was extracted using a DNA extraction kit and then after Real-Time PCR amplification, HRM was done.
Overall, 59 and 34 isolates were identified as
and
, respectively. The percentages of each species from animals were as follows: sheep (
, 80.39% and
, 19.61%), cattle (
, 42.85% and
, 57.15%).
HRM technique developed in the present study is a powerful, rapid and sensitive technique for epidemiological survey and molecular identification between
and
.
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