Occurrence of aflatoxin M1 in cheese products commonly available in Sri Lankan market.

Aflatoxins (AFs) are a group of mycotoxins produced by certain fungi of Aspergillus spp. AFs of major concern are B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2), and M1 (AFM1). AFM1 is a hydroxylated metabolite of AFB1 formed inside the animal's body which is excreted into milk of cows that consumed AFB1 contaminated feed. Consumption of AFM1-contaminated milk and subsequent dairy products causes negative health effects in consumers. This study was conducted to determine the occurrence and levels of AFM1 in cheese products available in the Sri Lankan market where AFM1 is not regularly monitored in milk while having an outdated regulatory limit of 1 ppb established for dairy products. Processed cheese (n = 28), hard cheese (n = 14), semi-hard cheese (n = 5), and soft cheese (n = 3) representing seven popular brands were collected. The samples were analyzed by Ultra High-Performance Liquid Chromatography Fluorescence Detection. AFM1 was detected in 40 samples (80 %), while 17 (34 %) and 37 (74 %) of the samples had AFM1 levels exceeding the maximum permitted limit set by Codex Alimentarius Commission (0.5 ppb) and the Netherlands (0.2 ppb). Further, 10 samples violated the Sri Lankan maximum limit of 1 ppb. Thirteen out of the 14 hard cheese (92.9 %, 0.11-14.43 ppb) and all semi-hard cheese samples (100 %, 0.29-0.65 ppb) contained AFM1. Most of the soft (66.7 %, 0.35-0.45 ppb) and processed (71.4 %, 0.11-1.35 ppb) cheese samples had AFM1. Most of the locally manufactured cheese products in Sri Lankan market may pose health risks to consumers. The results highlight the significance of regular monitoring of AFM1 in dairy products and the importance of updating regulations on par with international standards concurrently to ensure consumer safety.

Stable Isotope and Element Profiling for Determining the Agroclimatic Origin of Cow Milk within a Tropical Country.

Information on the geographic origin of milk is important in determining quality attributes and for economic gain through building brand value associated with origin. Stable isotope signatures and trace element concentrations have been increasingly used in authentication of milk, though information on the power of such technology in verifying agroclimatic origin in small continents with diverse climatic, environmental conditions, and animal management practice is scarce. Therefore, the main objective of this study was to investigate the possibility of using a stable isotope composition of C, N, O, and H and element fingerprints to determine the agroclimatic origin of milk produced in different agroclimatic zones of Sri Lanka. Stable isotopes ratios of C, N, H, and O, and elemental fingerprints of milk samples were determined by IRMS and ICP-MS, respectively. Significant variations were observed in stable isotope ratios, especially δ18O and the mean content of Li, Al, Cr, Mn, and Sr in the bulk milk samples obtained from different agroclimatic zones. A linear discriminant analysis differentiated cow milk produced from four agroclimatic zones based on stable isotope ratios, and the inclusion of elemental ratios enhanced the discriminating ability.

Carrier Status of Leptospira spp. in Healthy Companion Dogs in Sri Lanka.

Dogs are asymptomatic chronic carriers of Leptospira spp. and excrete these bacteria in their urine, resulting in environmental contamination and potentially leading to zoonotic transmission. Although a previous study in Sri Lanka detected anti-Leptospira antibodies in companion dogs, the urinary shedding of Leptospira spp. and the Leptospira species and serogroups prevalent in them remain unclear. Thus, the current study identified the prevalent Leptospira serogroups and the carrier status of Leptospira spp. in apparently healthy, client-owned dogs in the Kandy District of Sri Lanka. Serum and urine samples were collected from 96 unvaccinated and 82 vaccinated dogs. Anti-Leptospira antibodies and Leptospira DNA in urine were detected using the microscopic agglutination test (MAT) and nested PCR that targeted the pathogenic leptospiral gene, flaB. The flaB sequences were compared with those of Leptospira spp. using the public databases. MAT detected anti-leptospiral antibodies in 15.6% (15/96) of the unvaccinated dogs, and the reactive serogroups were observed to be Sejroe (11.5%), Canicola (2.1%), Icterohemorrhagiae (1.0%), and Javanica (1.0%). Furthermore, MAT results revealed that 11.0% (9/82) of the vaccinated dogs tested positive for the anti leptospira antibodies and the only reactive serogroup was Sejroe. Leptospira DNA was detected in 15.6% (15/96) and 15.9% (13/82) of urine samples collected from unvaccinated and vaccinated dogs, respectively, and phylogenetic analysis revealed that the animals were infected with L. borgpetersenii, L. interrogans, L. kmetyi, and L. weilii. The L. interrogans sequence detected in the canine sample was identical to the one that was previously reported in a human sample from the Kandy District. This study demonstrated that both unvaccinated and vaccinated dogs excrete various pathogenic Leptospira spp. in their urine, suggesting that they may play an important role in environmental contamination that poses a health risk to the dog owners and the general public.

Comprehensive Assessment of Essential and Potentially Toxic Trace Elements in Bovine Milk and Their Feeds in Different Agro-climatic Zones of Sri Lanka.

Fresh milk is an important source of essential mineral supplement for humans. However, the levels of trace elements in milk are an important component of its safety and quality. Trace elements also act as a bio-indicator of agricultural pollution. Samples of raw cow milk (n = 68), animal feed [forage (n = 36) and concentrates (n = 14)], and water (n = 35) were collected from different agro-climatic regions of Sri Lanka. The concentrations of 15 trace elements including toxic heavy metals such as Cd, Pb, and As were quantified using inductively coupled plasma mass spectrometry after microwave-assisted digestion. Among the studied trace elements, the mean elemental concentrations of Se, Cd, As, and Cu in cow milk were 18.1, 1.45, 7.35, and 71.7 µg L-1, respectively. The mean concentrations of these elements in forage were 0.216, 0.066, 0.046, and 9.21 mg kg-1, and in concentrate feed were 0.329, 0.202, 0.229, and 2.28 mg kg-1, respectively. The As content of the studied milk was well below the maximum permissible level while 15% of the samples had Pb exceeding the permissible limit of 20 µg L-1. However, As, Cd, and Pb levels in drinking water provided to animals were well below the WHO permissible limits. The results of this study revealed that the trace elements in cow milk depend mainly on the feed, and hence, levels of these contaminants in feed materials should be monitored. Although due to the consumption of milk, the populations have a minor exposure to trace elements and also minimum health risks, a detailed study with a large number of samples is highly recommended.

Tumor Spheroids as an in vitro model for determining the therapeutic response to proton beam radiotherapy and thermally sensitive nanocarriers.

Multicellular Tumor Spheroids (MCTS) strongly resemble tumor tissues, which makes them useful tools for radiation biology studies and screening of various chemotherapeutics. The goal of this pilot study was to use MCTS as an in vitro model to determine the response of cells to low temperature-sensitive liposomes (LTSLs) encapsulating doxorubicin (Dox) and proton beam radiotherapy (PBRT). Prior to treatment, MCTS were characterized for morphology and LTSLs were characterized for size, encapsulation efficiency, and ability to thermally release Dox (a model anticancer agent). Two groups of MCTS were treated with LTSL in combination with mild hyperthermia (40-42 °C) or PBRT alone in the presence of appropriate controls. Cytotoxic response was assessed after 48-72 h using an acid phosphatase assay. At 72 h, LTSL in combination with heat significantly reduced the viability of MCTS (15-30%) compared to the control (P < 0.05). A similar cytotoxic response was observed with PBRT treatment. The data suggest that like a monolayer cell culture, MCTS can be used to determine cytotoxic outcomes of thermal and proton therapy.

MRI-guided monitoring of thermal dose and targeted drug delivery for cancer therapy.

Application of localized hyperthermia treatment for solid tumor therapy is under active clinical investigation. The success of this treatment methodology, whether for tumor ablation or drug delivery, requires accurate target localization and real-time temperature mapping of the targeted region. Magnetic Resonance Imaging (MRI) can monitor temperature elevations in tissues in real-time during tumor therapy. MRI can also be applied in concert with methods such as High Intensity Focused Ultrasound (HIFU) to enable image-guided drug delivery (IGDD) from temperature sensitive nanocarriers, by exploiting not only its anatomic resolution, but its ability to detect and measure drug release using markers co-loaded with drugs within the nanocarriers. We review this rapidly emerging technology, providing an overview of MRI-guided tissue thermal dose monitoring for HIFU and Laser therapy, its role in targeted drug delivery and its future potential for clinical translation.

Canine intestinal histoplasmosis containing hyphal forms.

A 12-year-old intact male Miniature Schnauzer dog with chronic diarrhea that was unresponsive to empirical treatment was presented to a referring veterinarian. A laparotomy was performed, and formalin-fixed biopsies of duodenum, jejunum, and colon were sent to Oklahoma Animal Disease Diagnostic Laboratory for evaluation. Histologic examination revealed a severe, diffuse, granulomatous enteritis and colitis with intralesional yeast and hyphal forms. Grocott methenamine silver stains revealed short, aseptate hyphae co-mingled with 2-8 µm, oval to round yeast organisms consistent with Histoplasma capsulatum. The atypical presentation of both yeast and hyphal forms prompted identification of the organism. Direct sequencing of a polymerase chain reaction product from paraffin-embedded intestinal samples confirmed the presence of Ajellomyces capsulatus with a homology over 99% to several sequences in GenBank. Ajellomyces capsulatus is the holomorphic name for H. capsulatum. Therefore, the mycelial form of a dimorphic fungus such as H. capsulatum can coexist with yeast cells within lesions of histoplasmosis. Following diagnosis, the dog was treated with itraconazole for 6 months and has improved.