Previous research from our group reported that consumption of hamburger meat with spice mix added prior to cooking resulted in a reduction in the concentration of malondialdehyde, a lipid peroxidation marker, in the meat and in plasma and urine of healthy volunteers, and improved postprandial endothelial dysfunction in men with Type 2 diabetes . Subsequent study reported that commercial spices in dry or fresh form exhibited significant antioxidant capacity that correlated with total phenolic content butnot with the concentration of chemical biomarker . There is limited amount of information regarding the activity of culinary spice extracts against clinical isolated intestinal bacteria, and a limited number of bacterial strains have been assessed for their susceptibility or antimicrobial activity against spices. Gunes and colleagues reported that minimum inhibitory concentration of curcumin against 7 standard bacterial strains is in the range of 129 to 293 µg/mL . Cinnamaldehyde, a bio-active component of cinnamon, was shown to exhibit more potent in vitro antibacterial properties against 5 common foodborne pathogenic bacteria with MIC being 125 to 500 µg/mL as compared to crude cinnamon stick extract , but cinnamaldehyde did not modulate the population of selected Lactobacillus and Bifidobacterium counts in mouse cecal content . Supplementation of rosemary extract was reported to increase Bacteroides/Prevotella groups and reduce the Lactobacillus/Leuconostoc/Pediococcus group in the caecum of both obese and lean rats . Based on potential health benefits demonstrated from our group, this study investigated major chemical constituents, antioxidant activity, and in vitro effect of 7 spice extracts on the growth of 33 beneficial Bifidobacterium spp. and Lactobacillus spp.,square plastic pot and established their antimicrobial activity against 88 intestinal, pathogenic, and toxigenic bacterial strains.
Eighty-eight strains of bacteria included in this study were clinical isolates from patients in the Greater Los Angeles VA Healthcare Center as published , or purchased from American Type Culture Collection . Altogether 17 strains of Bifidobacterium species consisting of 11 strains recovered from human intestinal contents and 6 ATCC Bifidobacterium type strains were studied. Similarly, 16 strains of Lactobacillus species consisting of 12 strains recovered from human intestinal contents and 4 ATCC Lactobacillus type strains were studied. Other bacterialstrains, including 1 Akkermansia, Bacteroides spp., Clostridium spp., including 12 toxigenic Clostridium difficile, E. coli, Finegoldia magna, Fusobacterium spp., Peptoniphilus asaccharolyticus, Peptostreptococcus anaerobius, Ruminococcus spp., Salmonella typhi, and S. aureus strains, were also tested. These bacteria have been identified by 16S rRNA sequence analysis as published . Brucella agar plates containing serial dilutions of each spice extract and control plates without the tested spice were prepared . Spice extracts were 1st diluted by 20 times to give 9.0 mg/mL concentration which was further diluted to give 4.50, 2.25, 1.13, 0.56, 0.28, 0.14, and 0.07 mg/mL of extracts. A suspension of colonies was used to prepare Brucella broth tubes with a density equal to that of a 0.5 McFarland standard corresponding to density of 108 bacterial colony forming units /mL. A volume of 10 µL of these suspensions was inoculated on the spice extract-containing plates, achieving a final inoculum of 105 CFU/spot. Brucella agar plates were used as a positive growth control. The plates were incubated in an anaerobic chamber for 48 h at 37 °C. Anaerobic conditions consisted of a gas mixture of 5% CO2, 5% H2, and 90% N2; the residual oxygen was removed by palladium catalysts . After incubation, the plates were examined. The MICs were determined. MIC is defined as the lowest concentration of each spice resulting in no growth or a marked change in the appearance of growth as compared to the control plate, as described in the Clinical and Laboratory Standards Institute approved protocol .
Further, the concentrations resulting in the change of the appearance of growth as compared to the control plate were recorded to establish for each test strain the concentration where the tested spice had no effect on the growth, where the spice stimulated the growth, and where the growth stimulation by the test spice reached a plateau .Results from our chemical investigations indicated that most spice extracts contain either moderate or high content of spice specific phytochemicals. The identification and quantitation of these phytochemicals provide insight into the potential influence of these chemicals on the gut microbial communities and activities. Among the chemicals characterized, majority belongs to polyphenols and phenolic acids. Convincing evidences suggest that the beneficial effects attributable to dietary phenols depend on their biotransformation by the gut microbiota. Most polyphenols present in the diet are in the form of esters, glycosides, or polymers that cannot be absorbed in their native form. It is estimated that about 5% to 10% of dietary polyphenols are absorbed and reach plasma. Over 95% of the intake passes to the colon and is fermented by gut microbiota to produce small aromatic or phenolic acids . For example, rosmarinic acid, found abundantly in ORE, ROS, and in many Lamiaceae plants, is an ester of caffeic acid and 3-lactic acid. A recent study reported a complete degradation of rosmarinic acid, and the generation of microbial product caffeic acid after in vitro fermentation using human feces . The caffeic acid undergoes subsequent microbial transformation to yield hydroxyphenylpropionic acids as major metabolites, a process mediated by microbial chlorogenate esterases or by Lactobacillus johnsonii . Results of human intervention study confirmed these in vitro findings . These and other microbial metabolites were found to selectively inhibit the growth of pathogenic bacteria and stimulate the growth of beneficial microorganisms .Of the 16 Lactobacillus strains tested, the growth of 1 strain was enhanced by BLP and another one by CAP at the concentration of 0.56 mg/mL. At 1.13 mg/mL, 4 strains were stimulated by BLP, 5 by CAP, 7 by GIN, and 10 by ORE. At 2.25 mg/mL, 2 strains were stimulated by BLP, 7 by CAP, 4 by GIN, and 2 by TUR. Overall, all spice extracts showed better growth stimulatory effect on the Lactobacillus spp. as compared to Bifidobacterium spp. Some foods can exert growth stimulatory effect because their ingredients can be used as substrates by these bacteria, or they can enhance nutrients consumption by affecting bacteria metabolism .
Prebiotics are defined as substances that induce the growth or activity of microorganisms that contribute to the well-being of their host . Lactobacillus and Bifidobacterium are widely established bacterial genera as prebiotic target organisms. These 2 genera produce acetic acid and lactic acid as the major end-metabolites and do not contain any known pathogens. Previous studies by our group and others have identified dietary sources functioning as prebiotics by stimulating the growth of Bifidobacterium spp. and Lactobacillus spp. in both in vitro and human studies . Evidence is growing in support of the prebiotic effect of foods high in polyphenols. Our results suggest that some of the tested spices exhibit prebiotic-like effect by stimulating the growth of a number of Bifidobacterium and Lactobacillus species. Bifidobacteria are normal inhabitants of the gastrointestinal tract. The composition, diversity, or relative abundance of Bifi- dobacterium species has been implicated in several intestinal disease conditions . Bifidobacteria have been shown to alleviate infectious diarrhea through their effects on the immune system . Lower levels of bi- fidobacteria were linked to higher prevalence of E. coli in obese children ; similarly,square pot lower levels of bifidobacteria have been demonstrated in overweight subjects and in patients with long-term asthma , suggesting a role of bifidobacteria in human health. Our study reported new findings in that among 7 spice extracts ORE was most active in promoting the growth of Bifi- dobacterium whereas GIN, BLP, and CAP produced more modest stimulatory activity. Studies of Lactobacillus rhamnosus GG have shown promising results in treating diarrhea caused by viruses and bacteria , atopic disease , and in prevention of gastrointestinal and respiratory tract infections . In our study, all spice extracts were found to promote the growth of LGG strain at concentration ranging from 0.56 to 4.5 mg/mL. L. reuteri and L. rhamnosus are often added to dairy products, or formulated as dietary supplements for controlling dysbiotic bacterial overgrowth during an active infection. BLP, CAP, GIN, and ORE enhanced the growth of these species at either 1.13 or 2.25 mg/mL. Our data are consistent with a study by Sutherland who reported that aqueous extracts of ginger and red chili enhanced the growth of L. reuteri and L. rhamnosus .Both medical students and faculty recognized the need for accessible health care among marginalized populations in the Zona Norte, and in 2011, a bi-national student-run free clinic was established in the heart of Tijuana’s red-light district . The clinic known as HFiT, or Health Frontiers in Tijuana, partners with Tijuana’s Autonomous University of Baja California School of Medicine and the University of California, San Diego School of Medicine. HFiT is equipped to provide medication, basic laboratory tests, and social services to patients living in poverty. Despite the clinic’s success in providing continuity care for more than 600 patients in 2012, the HFiT clinical faculty and student leadership recognized the need for a vulnerable women’s health clinic. An HFiT survey conducted between May and December 2012, found that of the 211 female patients, 12% stated they required obstetrical or gynecological care. Apart from HIV and syphilis diagnostic testing, the HFiT clinic currently lacks the supplies to perform other sexually transmitted infection testing, pap smears, endometrial biopsies, or provide contraception.
Furthermore, increasing HIV prevalence among female sex workers in Tijuana emphasizes the need for interventions dedicated to the education and prevention of HIV transmission. In a study of 924 FSWs, nearly one third of all questions on HIV transmission and prevention mechanisms were answered incorrectly . Therefore educating and refuting common HIV misconceptions among FSWs demonstrates a potentially powerful intervention method for HIV prevention. The HFiT clinic has provided the physical space for a women’s clinic. This includes a private exam room and an exam table. Our plan of action is geared towards formally establishing a Women’s Clinic that will address the needs of our vulnerable patient population and will sustain long-term contribution to the clinic.Beginning the summer of 2013, after each Saturday clinic two clinic volunteers and I went around Tijuana’s Zona Roja in order to distribute flyers advertising our services and clinic location. We also distributed condoms and handouts on how to properly use a condom. Although we handed out maps/flyers to females all around the clinic, we tried to target the areas where most sex work took place in the Zona Roja. Here we asked female sex workers if they were willing to fill out a brief, anonymous survey asking where they currently go for medical attention, what medical services they need, and whether they were aware of our HFiT clinic. The surveys evolved over the course of the summer as we found some questions to be more effective than others and many FSWs did not have more than 5 minutes to devote to answering survey questions. Although responses varied for desired medical services, based on the results most sex workers did not know about our clinic prior to taking the survey. With a finalized version of the survey currently in place, we will continue to assess the population and evaluate how responses change over time. After researching the most cost-effective options for an ultrasound, including contacting the Professor of Clinical Emergency Medicine and Director of Instructional Ultrasound at UC Irvine School of Medicine, we found that our past contact for much of the clinic supplies was the best and most reliable option. We were able to secure an ultrasound complete with wide-angle probe, vaginal probe, and printer for a total of $2300. The ultrasound is refurbished, recalibrated and in great condition. Although the budget for the summer did not allow for purchase of both ultrasound and colposcope, we are very pleased that the ultrasound with endoprobe will allow us to perform transvaginal ultrasounds to assess gynecological anatomy.Following the clinic’s goal of empowering its patients, we have designed a Women’s Health curriculum that explores female anatomy, body perception, and general nutrition. The implementation of this program and its success can be attributed to the collaboration with Por La Vida . Por La Vida began as a pilot health education-advising program in San Diego in 1987. Evidence of its success and longevity 25 years later is seen through the over 200 “Consejeras” or advisors that currently lead small group sessions in the medically under served Latino Community of San Diego.