Development of novel isothermal-based DNA amplification assay for detection of pig tissues in adulterated meat.

For the first time, we describe an innovative polymerase spiral reaction (PSR) assay for the rapid, simple, and accurate detection of pig tissues or pork in adulterated meat including heat-treated and processed ones. The PSR assay specifically targeting the mitochondrial cytochrome b (cyt-b) gene of the pig was successfully optimized permitting assay results in 65 min time. The developed detection method was 100% specific amplifying only the cyt-b gene and displaying negative results with all the tested non-pork meats. The sensitivity of the developed PSR (760 fg porcine DNA) was tenfold better than the end-point PCR and able to detect heat-treated (121 °C) and adulterated (0.5% pork in beef) meat and processed pork products such as sausages, salami, meatball, soup, curry, etc. The developed PSR-based method can be used for point-of-care detection with minimum instrumentation and technical expertise to guarantee instant clearance of exported and imported meat products. This is the first time that PSR has been adapted for food authenticity purposes.

Isolation, susceptibility profiles and genomic analysis of a colistin-resistant Salmonella enterica serovar Kentucky strain COL-R.

Salmonella enterica serovar Kentucky is a frequent cause for clinical infections in human patients. They are isolated and reported with multidrug resistance from the foods of animal origin from various countries. However, studies inferring the colistin resistance are limited. Hence, the current study reports the genetic factors and genomic analysis of the colistin-resistant Salmonella enterica serovar Kentucky strain COL-R for better understanding of its pathogenic potential and phylogenetic relatedness. The S. Kentucky strain COL-R was successfully isolated from chicken meat during ongoing surveillance of food of animal origin. Antimicrobial susceptibility testing revealed resistance to cefoxitin, erythromycin, gentamicin, tetracycline, and most disturbingly to ciprofloxacin and colistin (broth microdilution method). Whole-genome sequence of the COL-R strain was subjected to various in silico analysis to identify the virulence factors, antimicrobial resistance genes, pathogenicity islands and sequence type. The S. Kentucky COL-R strain belonged to sequence type (ST) 198 with a high probability (0.943) of being a human pathogen. Besides presence of integrated phage in the S. Kentucky COL-R genome, 38 genes conferring resistance to various antimicrobials and disinfectants were also identified. Nucleotide Polymorphism analysis indicated triple mutations in gyrA and parC genes conferring fluoroquinolone resistance. Phylogenomic analysis with 31 other S. Kentucky genomes revealed discernible clusters with S. Kentucky COL-R strain latching onto a cluster of high diversity (geographic location and isolation sources). Taken together, our results document the first occurrence of colistin resistance in a fluoroquinolone resistant S. Kentucky COL-R strain isolated from retail chicken and provide crucial information on the genomic features of the strain. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03559-2.

Salmonella from Farm to Table: Isolation, Characterization, and Antimicrobial Resistance of Salmonella from Commercial Broiler Supply Chain and Its Environment.

Antimicrobial resistance (AMR) in poultry production chain is one of the major food safety concerns due to indiscriminate usage of antibiotics and the presence of pathogens such as Salmonella which causes infections in various stages of production. In the present study, 182 samples were collected from commercial broiler supply chain, viz., three hatcheries (n = 29), three commercial broiler farms (CBF; n = 99), and three retail meat shops (RMS; n = 54), and used for isolation and identification of Salmonella using three different selective agar media and a selective enrichment medium followed by PCR confirmation targeting the hilA gene. The overall prevalence of Salmonella was 47/182 (25.82%), and a significantly higher (P < 0.05) prevalence was observed in retail meat shops (46.29%), CBF (19.19%), and hatcheries (10.34%). Comparison of three agar media for isolation of Salmonella revealed that all the media were equally selective. However, PCR amplification of hilA gene fragment was significantly higher (P < 0.01) in selective enrichment culture tetrathionate brilliant green bile broth (TTB) as compared to all solid (agar-based) media. Susceptibility pattern against most frequently used antibiotics revealed that 100% of the isolates were resistant to at least one antibiotic. High resistance was observed for doxycycline (94.34%), followed by cefpodoxime (84.91%), ciprofloxacin (72.64%), gentamicin (65.09%), enrofloxacin (61.32%), colistin sulphate (40.42%), amikacin (34.91%), ampicillin (33.96%), neomycin (33.02), cefotaxime (30.19%), ceftazidime (29.25%), trimethoprim-sulfamethoxazole (23.58%), amoxicillin+clavulanic acid (21.70%), and chloramphenicol (12.26%); 16.98% of the isolates were ex-tended spectrum ß-lactamase (ESBL) producers, and 76.41% were multidrug resistant (MDR). MDR Salmonella were significantly higher (P < 0.01) in RMS (91.66%) followed by CBF (82.75%), whereas no MDR isolates were present in the isolates from hatcheries. The results indicated a higher prevalence of Salmonella and AMR for commonly used antibiotics in the complete broiler supply chain, especially RMS and CBF. Also, this study idicated that TTB enrichment followed by PCR and colony PCR was found to be rapid, specific and time-saving method.

Anticancer efficacy of 6-thioguanine loaded chitosan nanoparticles with or without curcumin.

6-Thioguanine encapsulated chitosan nanoparticles (6-TG-CNPs) has formulated by the ionic-gelation method. Morphologically, the 6-TG-CNPs were spherical and showed mean size, PDI, zeta potential, and entrapment efficiency of 261.63 ± 6.01 nm, 0.34 ± 0.10, +15.97 ± 0.46 mV and 44.27%, respectively. The IR spectra confirmed the 6-TG complex with chitosan. The in vitro drug release profile of 6-TG-CNPs revealed an increase in sustained-release (91.40 ± 1.08% at 48 h) at pH 4.8 compared to less sustained-release (73.96 ± 1.12% at 48 h) at pH 7.4. The MTT assay was conducted on MCF-7 and PA-1 cell lines at 48 h incubation to determine % cell viability. The IC50 values of 6-TG, 6-TG-CNPs, and curcumin for MCF-7 were 23.09, 17.82, and 15.73 µM, respectively. Likewise, IC50 values of 6-TG, 6-TG-CNPs, and curcumin for PA-1 were 5.81, 3.92, and 12.89 µM, respectively. A combination of 6-TG-CNPs (IC25) with curcumin (IC25) on PA-1 and MCF-7 showed % cell viability of 43.67 ± 0.02 and 49.77 ± 0.05, respectively. The in vitro cytotoxicity potential in terms of % cell viability, early apoptosis, G2/M phase arrest, and DNA demethylating activity of 6-TG-CNPs alone and combination with curcumin proved to be more effective than that of 6-TG on PA-1 cells.

TB care for women and Covid-A double health crisis in the offing?

The COVID-19 pandemic and its health, economic and social aftermath has currently overshadowed every other health concern throughout the world. There are numerous ways in which this will impact existing public health issues. In this article we aim to examine and reflect on the interactions between COVID-19 and tuberculosis (TB) another scourge which has long been the world's leading infectious killer. There are potentially grave consequences for existing and undiagnosed TB patients globally, particularly in low and middle income countries like India where TB is endemic and health services poorly equipped. TB control programmes are likely to be strained due to diversion of resources, and an inevitable loss of health system focus. This is likely to lead to a reduction in quality of TB care and worse outcomes. The symptoms of TB and COVID-19 can be similar, for instance symptoms like cough and fever. Not only can this create diagnostic confusion, but it could worsen the stigmatization of TB patients especially in LMICs, given the fear of COVID-19. Women with TB in India are a vulnerable group as gender disparities play a significant role in how men and women access healthcare in the public and private sectors. They are especially likely to suffer as part of the "collateral damage" and also experience the impact of the double stigma disproportionately. In this paper the researchers examine the impact of Covid 19 on women seeking TB care in India.

Understanding patient choices regarding breast reconstruction after mastectomy for breast cancer.

PURPOSE: In Australia, about 40% of patients undergo mastectomy to treat breast cancer, with negative impacts on body image, sexual function and quality of life. Whilst breast reconstruction is associated with increased patient self-esteem and a greater sense of wholeness and well-being, the national reconstruction rate is low at 18%. This study aimed to compare demographics, treatment factors and information provision about breast reconstruction in women who had and did not have breast reconstruction following mastectomy treatment and identify goals and concerns underpinning women's reconstruction decisions. METHODS: Female patients who had a mastectomy to treat breast cancer between 2010 and 2014 in a culturally and linguistically diverse (CALD) and socially disadvantaged region participated in a cross-sectional study, completing a questionnaire in their language of choice (English, Vietnamese, Chinese or Arabic). RESULTS: Completed surveys were returned by 168 women (42% response rate; 77% English-speaking), of whom only 19.0% (n = 32) reported having had breast reconstruction. Reconstruction rates were significantly lower in women who reported speaking a language other than English at home versus only English (37.5% vs 62.5%, p = 0.03). However, all women expressed a desire for more information about breast reconstruction and more support to make their decision about breast reconstruction. CONCLUSIONS: Patients identified a need for greater information provision on breast reconstruction, highlighting an urgent need for resources specifically about breast reconstruction, particularly for non-English-speaking patients. Greater provision of information prior to mastectomy is critical to underpin breast cancer patients' decisions about breast reconstruction, especially for non-English speaking patients.

[Comparison CCR5de132 mutation in the CCR5 gene frequencies in Russians, Tuvinians, and in different groups of HIV-infected individuals].

The 32-bp deletion (CCR5del32 mutation) in the CCR5 (chemokine (C-C motif) receptor 5) gene, encoding CCR5 chemokine receptor, is one of the factors determining natural resistance to human immunodeficiency virus (HIV-1) infection. In the present study, the samples of Russians (n = 107), Tuvinians (n = 50), and HIV-infected individuals were examined for the presence of CCR5del32 mutation in the CCR5 gene. The CCR5del32 allele frequency in Russians and Tuvinians constituted 7.84 and 2%, respectively. Among HIV-1 infected individuals, two groups, of macrophage-tropic HIV-1 strain- and T-cell-tropic HIV-1 strain-infected were distinguished. The CCR5del32 allele frequency in the first group (6.45%) was lower than in the second one (8.73%). Statistical treatment of the HIV-1 infected individuals typing data showed that the difference in the CCR5del32 allele frequencies between the groups of sexually (macrophage-tropic) and parenterally (T-cell-tropic) infected individuals observed was within the limit of random deviation.

[Rapid diagnosis of dysbacteriosis by detecting beta-aspartylglycine in the feces in an experiment]. / Ekspressnaia diagnostika disbakterioza putem obnaruzheniia beta-aspartil-glitsina v fekaliiakh v éksperimente.

In experiments on Fischer rats (F-344), both with common microflora and germ-free, the influence of the systemic destination of different antibiotics (tetracycline + ampicillin, gentamicin + kefzol, gentamicin, fradizine) on the intestinal microflora, the content of beta-ospartylglycine in feces and the colonization resistance of the intestinal tract to Staphylococcus aureus B-243 and Pseudomonas aeruginosa No. 93 has been studied. The early appearance of beta-aspartyl-glycine in the supernatant of fecal samples has been shown to be the first sign of dysbacteriosis and to indicate the decrease of the colonization resistance of the intestine to opportunistic bacteria.

[Immunoglobulins A and G in the intestinal secretions of rats in different microbiologic conditions]. / Immunoglobuliny A i G v sekrete kishechnika krys pri razlichnom mikrobiologicheskom statuse.

Experiments were made with 3 groups of inbred CDF (F344) Crl rats: 30 germ-free, 30 conventional and 18 germ-free animals infected orally with Shigella flexneri 2aN 516. Secretory IgA (S-IgA) was isolated from gut secretion of the conventional rats to obtain rabbit antiserum against it. The levels of S-IgA, IgA and IgG in blood serum and gut secretion were determined. It was shown that gut secretion of the germ-free rats contained no less S-IgA than that from the conventional rats, at the same time IgG was not found in the germ-free rats. After injection of the germ-free rats with Shigella flexneri the content of S-IgA in gut secretion did not rise but there appeared IgG which differed immunochemically from serum IgG2. Within the first week after the injection, gut secretion showed specific antibodies to Shigella flexneri, which was ascertained with the aid of the hemagglutination inhibition test. The titers of antibodies exceeded 1:4096 by day 21. Antibodies in serum appeared much later and in low titers (1:16, 1:32). Therefore, the local immune response was marked to a greater degree than the generalized one. In spite of the presence of specific antibodies in high titers, Shigella flexneri persisted in the hosts, i. e. one could observe a clinically normal bacterial carriage.

[Action of cyclophosphane on the formation of hemagglutinins and of nonspecific immunoglobulins in a microbe-free state]. / Deistvie tsiklofosfana na obrazovanie gemaggliutininov i nespetsificheskikh immunoglobulinov pri stause bezmikrobnosti.

A total of 39 germ-free and 28 conventional germ-free Fisher rats were used to study cyclophosphamide-induced specific (hemagglutinins) and nonspecific (IgG2) immunosuppression. Most pronounced effect of the drug was observed in germ-free animals. 180 mg/kg of cyclophosphamide completely suppressed hemagglutinin formation for at least 8 weeks.

[Effect of normal microflora, age and sex of rats on serum IgG2 levels]. / Vliianie normal'noi mikroflory, vozrasta i pola krys na uroven' IgG2 v syvorotke krovi.

Germfree and conventional rats of Fisher inbred strain and mongrel animals were used in this study. IgG2 was separated from the pool serum of mongrel rats, and anti-IgG2 was obtained by hyperimmunization of rabbits. IgG2 content was determined by in the sera of different groups of rats radial immunodiffusion. There was 5.4 times less IgG2 in the sera of Fisher germfree rats than in conventional Fisher animals; its level in female animals was by 10--15% higher than in the male ones. The first hours after birth the IgG2 level was the same in the sera of all the newborn rats irrespective of its amount in the maternal sera.