Philipus Pangloli

Prevalence of Salmonella in Diverse Environmental Farm Samples

The development of suitable intervention strategies to control Salmonella populations at the farm level requires reliable data on the occurrence and prevalence of the pathogen. Previous studies on Salmonella prevalence have focused on acquiring data from specific farm types and/or selected regions. The purpose of this study was to evaluate the distribution of this pathogen across a variety of farm types and regions in order to generate comparative data from a diverse group of environmental samples. Farm samples (n = 2,496) were collected quarterly from 18 different farms across five states (Tennessee, North Carolina, Alabama, California, and Washington) over a 24-month period. The participating farms included beef and dairy cattle operations, swine production and farrowing facilities, and poultry farms (both broiler chicken and turkey). The samples were analyzed for the presence of Salmonella by means of the U.S. Food and Drug Administrations Bacteriological Analytical Manual methods optimized for farm samples. Salmonella isolates were characterized by automated riboprinting. Salmonella serovars were recovered from 4.7% of all samples. The majority of positive findings were isolated from swine farms (57.3%). The occurrence of Salmonella was lower on dairy farms (17.9%), poultry farms (16.2%), and beef cattle farms (8.5%). The most commonly isolated serovar was Salmonella Anatum (48.4%), which was isolated notably more frequently than the next most common Salmonella serovars, Arizonae (12.1%) and Javiana (8.8%). The results of this study suggest that significant reservoirs of Salmonella populations still exist on swine production facilities and to a lesser extent in other animal production facilities. Data showed that the surrounding farm environment could be an important source of contamination.

Occurrence of Escherichia coli O157:H7 in diverse farm environments

In the United States, foodborne outbreaks of Escherichia coli O157:H7 illness have often been linked to the consumption of contaminated, undercooked ground beef. However, the occurrence of E. coli O157:H7 has also been reported in other farm animals. The objective of this study was to evaluate the occurrence of E. coli O157:H7 on diverse farm types and from a variety of farm samples. Rectal swabs (n=1686) and environmental samples (n=576) were collected from 16 farms in five states over 24 months and analyzed for the presence of E. coli O157:H7. Overall, E. coli O157:H7 was found in 3.6% of beef cattle, 3.4% of dairy cattle, 0.9% of chicken, 7.5% of turkey, and 8.9% of swine samples. The pathogen was isolated sporadically from each of the environmental sample types. Of particular concern was the isolation of E. coli O157:H7 from fresh feed samples, indicating a potential vector for transmission. The data from this study indicate a high occurrence of E. coli O157:H7 on swine and turkey farms. This unexpected result suggests that more research on the occurrence of E. coli O157:H7 on these types of farms is required in order to better understand potential reservoirs of pathogenic E. coli.

Seasonal incidence and molecular characterization of Salmonella from dairy cows, calves, and farm environment.

The occurrence patterns and molecular characteristics of Salmonella are important for surveillance and control of the pathogens. Objectives of this study were to determine month-to-month variation and seasonal effects on the occurrence of Salmonella in dairy animals and environments and to characterize selected Salmonella isolates. A total of 7680 animal and environmental samples, collected monthly from a dairy farm, were analyzed for the presence of Salmonella during a 12-month study. Major sources of Salmonella on the dairy farm (% positive) were milking parlor air (62%) and bird droppings (63%) during winter; feeds (50-58%), water (53-67%), calf bedding (63%), soils (60-63%), milking parlor air (60%), and bird droppings (50%) in spring; all animal and environmental samples (40-92%) except milking parlor air (25%) and bulk tank milk (29%) in summer; and feeds (60-71%), cow beddings (59%), cow soils (50%), air (46-71%), and insects (63%) during fall. Salmonella ribotyping indicated that most serovars came from different sources but some might have originated from a common source and transmitted from site to site on the farm. These data provide some important information on key animal and environmental sampling sites needed to initiate on-farm management programs for control of this important foodborne pathogen.

Evaluation of Methods for Recovery of Salmonella from Dairy Cattle, Poultry, and Swine Farms

Current official methods for detection and isolation of Salmonella are mostly designed for foods. The objective of this study was to determine optimal methods for detection and isolation of Salmonella from animal and environmental samples of dairy, poultry, and swine farms. Preenrichment in lactose broth versus direct enrichment (no preenrichment) prior to selective enrichment in Rappaport-Vassiliadis, selenite cystine, and tetrathionate incubated at 35 and 42 degrees C and in four differential/selective plating media (brilliant green, bismuth sulfite, Hektoen enteric, and xylose-lysine-tergitol 4 agar base) were evaluated for their ability to recover Salmonella from artificially contaminated samples. The effects of pH adjustments to samples on Salmonella recovery were determined. A pH adjustment of the enrichment broth to 6.8 +/- 0.2 after addition of samples significantly improved recovery of Salmonella. The most effective medium combinations for isolation of Salmonella from farm samples depended on the type of samples. Generalizations of protocols for recovery of Salmonella from farm samples might result in poor recovery, increased recovery time, and increased sample processing costs.

Epithelial-to-Mesenchymal Transition in Paclitaxel-Resistant Ovarian Cancer Cells Is Downregulated by Luteolin

Ovarian cancer (OVCA) is the deadliest of all gynecological cancers which is attributed to late presentation, persistence, and development of chemoresistance. The objectives were to evaluate the association between OVCA paclitaxel‐resistance and epithelial‐to‐mesenchymal transition (EMT) and to determine the capability of luteolin to chemosensitize OVCA cells. X10 and X22 cells were 11.8–25.3‐fold and 7.8–8.6‐fold resistant to paclitaxel than 1AP cells. X10 and X22 cells exhibited a mesenchymal phenotype, while 1AP has an epithelial characteristics. Furthermore, the expression of the epithelial marker E‐cadherin was downregulated, while mesenchymal markers Vimentin and N‐cadherin were upregulated in X10 and X22 cells when compared to 1AP cells. Transcription factors Snail, Slug, and Twist1 were upregulated in X10 cells, while Twist1 was highly expressed in X22 cells. Luteolin treatment caused cytotoxicity being most potent to X10 OVCA cells. Treatment of non‐cytotoxic dose of luteolin at 15.625 μM chemosensitized X10 and X22 OVCA cells to paclitaxel as evidenced by reduced ED50 values from 11.8 to 0.2 μM and 8.6 to 3.6 μM for X10 and X22 cells, respectively. Moreover, luteolin treatment led to a more epithelial phenotype of X10 and X22 cells and modification of EMT markers indicating reversal of EMT. The mechanism involved is through reduction of phosphorylation of FAK and ERK leading to reduced nuclear translocation of p65. Our results highlight the significance of EMT in OVCA resistance to paclitaxel and warrant the investigation of luteolin as a potential therapeutic agent in chemoresistant OVCA. J. Cell. Physiol. 232: 391–401, 2017.

Kunitz trypsin inhibitor in addition to Bowman-Birk inhibitor influence stability of lunasin against pepsin-pancreatin hydrolysis

Soybean contains several biologically active components and one of this belongs to the bioactive peptide group. The objectives of this study were to produce different lunasin-enriched preparations (LEP) and determine the effect of Bowman-Birk inhibitor (BBI) and Kunitz trypsin inhibitor (KTI) concentrations on the stability of lunasin against pepsin-pancreatin hydrolysis (PPH). In addition, the effect of KTI mutation on lunasin stability against PPH was determined. LEP were produced by calcium and pH precipitation methods of 30% aqueous ethanol extract from defatted soybean flour. LEP, lunasin-enriched commercially available products and KTI control and mutant flours underwent PPH and samples were taken after pepsin and pepsin-pancreatin hydrolysis. The concentrations of BBI, KTI, and lunasin all decreased after hydrolysis, but they had varying results. BBI concentration ranged from 167.5 to 655.8μg/g pre-hydrolysis and 171.5 to 250.1μg/g after hydrolysis. KTI concentrations ranged from 0.3 to 122.3μg/g pre-hydrolysis and 9.0 to 18.7μg/g after hydrolysis. Lunasin concentrations ranged from 8.5 to 71.0μg/g pre-hydrolysis and 4.0 to 13.2μg/g after hydrolysis. In all products tested, lunasin concentration after PPH significantly correlated with BBI and KTI concentrations. Mutation in two KTI isoforms led to a lower concentration of lunasin after PPH. This is the first report on the potential role of KTI in lunasin stability against PPH and must be considered in designing lunasin-enriched products that could potentially survive digestion after oral ingestion.

BG-4, a novel bioactive peptide from Momordica charantia, inhibits lipopolysaccharide-induced inflammation in THP-1 human macrophages

BACKGROUND Bitter melon (Momordica charantia) is a commonly used food crop for management of a variety of diseases most notably for control of diabetes, a disease associated with aberrant inflammation. PURPOSE To evaluate the anti-inflammatory property of BG-4, a novel bioactive peptide isolated from the seed of bitter melon. METHODS Differentiated THP-1 human macrophages were pre-treated with BG-4 and stimulated with lipopolysaccharide. Pro-inflammatory cytokines IL-6 and TNF-α were measured by enzyme-linked immunosorbent assay. The mechanism of action involving activation of NF-κB and phosphorylation of ERK and STAT3 was measured by western blot and immunofluorescence. The production of intracellular reactive oxygen species was evaluated by fluorescence microscopy and fluorescence spectrophotometry. RESULTS BG-4 dose dependently reduce the production of pro-inflammatory cytokines IL-6 and TNF-α. The ability of BG-4 to reduce production of cytokines are associated with reduced phosphorylation of ERK and STAT3 accompanied by reduced nuclear translocation of p65 NF-κB subunit. The mechanism of action is reduction of LPS-induced production of intracellular reactive oxygen species. CONCLUSION Our results demonstrated the ability of BG-4, a novel peptide from the seed of bitter melon, to exert anti-inflammatory action. This could explain the traditional use of bitter melon against diseases associated with aberrant and uncontrolled inflammation.

Impact of ultrasonication on the physicochemical properties of sorghum kafirin and in vitro pepsin-pancreatin digestibility of sorghum gluten-like flour

Sorghum is a nutritionally dense grain but its nutritional quality is limited by its poor digestibility. The objective was to determine the effects of ultrasonication on the physicochemical properties of kafirin, major protein in sorghum, and its stability against pepsin-pancreatin hydrolysis (PPH). Ultrasonication for 10min at 40% amplitude increased the solubility of purified kafirin from 6.5μg/mL to 173.3μg/mL. Ultrasonication altered the secondary structure of kafirin as evaluated by circular dichroism and Fourier-transform infrared red spectroscopy. In pepsin-pancreatin hydrolysates, 14.7% increase in molecules with molecular weight between 0.075 and 0.5kDa was detected in size-exclusion chromatogram after ultrasonication at 40% amplitude, 10min. The degree of hydrolysis was also increased after ultrasonication by 17.7%, 127.6%, 346.6% as measured by o-phthaldialdehyde derivatization, trichloroacetic acid precipitation and hydrochloric acid hydrolysis, respectively with improved antioxidant property. Our results showed the potential of ultrasonication to improve digestibility and biological properties of sorghum flour.