Baird-Parker Agar Research Articles

Analysis of Microbiological Hazards in Turmeric Tamarind Traditional Drink Products

Turmeric tamarind is a traditional drink with a sour and sweet taste traditionally made on a household scale. This drink is popular among the community, students, and local tourists from various regions visiting Yogyakarta. Due to its relatively simple production process and potentially suboptimal hygiene standards, the risk of bacterial contamination remains a concern for public health. Therefore, this study aimed to determine the presence or absence of bacterial contaminants, namely Escherichia coli, Salmonella sp and Staphylococcus aureus according to SNI 2019 standards. Samples were obtained from 11 locations of turmeric tamarind drink sales and inoculated into Chromocult Coliform Agar (CCA) differential selective medium, Salmonella Sigella Agar (SSA), and Baird Parker Agar (BPA) for E. coli isolation, Salmonella sp, and S. aureus, respectively. Subsequently, colonies were prified and screened through biochemical tests such as IMViC, Urease test, and sugar fermentation test for lactose, maltose, and glucose. Suspected isolates were also confirmed using API 20E and API STAPH tests. The results showed that E. coli, Salmonella sp, and S. aureus were not reported while Klebsiella pneumoniae ssp pneumoniae 1, Citrobacter freundii, Staphylococcus xylosus, and Staphylococcus lentus were obtained.

Occurrence, antimicrobial susceptibility, and resistance genes of Staphylococcus aureus in milk and milk products in the Arsi highlands of Ethiopia.

In Ethiopia, milk production and handling practices often lack proper hygiene measures, leading to the potential contamination of milk and milk products with Staphylococcus aureus (S. aureus), including methicillin-resistant strains,posing significant public health concerns. This study aimed to investigate the occurrence, antimicrobial susceptibility profiles and presence of resistance genes in S. aureus strains isolated from milk and milk products. A cross-sectional study was conducted in the Arsi highlands, Oromia, Ethiopia from March 2022 to February 2023. A total of 503 milk and milk product samples were collected, comprising 259 raw milk, 219 cottage cheese, and 25 traditional yogurt samples. S. aureus isolation and coagulase-positive staphylococci enumeration were performed using Baird-Parker agar supplemented with tellurite and egg yolk. S. aureus was further characterized based on colony morphology, Gram stain, mannitol fermentation, catalase test, and coagulase test. Phenotypic antimicrobial resistance was assessed using the Kirby-Bauer disc diffusion method, while the polymerase chain reaction (PCR) was employed for confirming the presence of S. aureus and detecting antimicrobial resistance genes. S. aureus was detected in 24.9% of the milk and milk products, with the highest occurrence in raw milk (40.9%), followed by yogurt (20%), and cottage cheese (6.4%). The geometric mean for coagulase-positive staphylococci counts in raw milk, yogurt, and cottage cheese was 4.6, 3.8, and 3.2 log10 CFU/mL, respectively. Antimicrobial resistance analysis revealed high levels of resistance to ampicillin (89.7%) and penicillin G (87.2%), with 71.8% of the isolates demonstrating multidrug resistance. Of the 16 S. aureus isolates analyzed using PCR, all were found to carry the nuc gene, with the mecA and blaZ genes detected in 50% of these isolates each. This study revealed the widespread distribution of S. aureus in milk and milk products in the Arsi highlands of Ethiopia. The isolates displayed high resistance to ampicillin and penicillin, with a concerning level of multidrug resistance. The detection of the mecA and blaZ genes in selected isolates is of particular concern, highlighting a potential public health hazard and posing a challenge to effective antimicrobial treatment. These findings highlight the urgent need to enhance hygiene standards in milk and milk product handling and promote the rational use of antimicrobial drugs. Provision of adequate training for all individuals involved in the dairy sector can help minimize contamination. These measures are crucial in addressing the threats posed by S. aureus, including methicillin-resistant strains, and ensuring the safety of milk and its products for consumers.

Prevalence, molecular identification and determination of antibiotic susceptibility of methicillin-resistant Staphylococcus aureus in raw meat

Objective: The aim of this study was to determine the prevalence, molecular identification, and antibiotic susceptibility of MRSA in raw meats of retail sale in Balikesir, Turkey. Materials and Methods: A total of 250raw meat samples (beef n=100, chicken n=100, and turkey n=50) were collected from various supermarkets. Mueller-Hinton Broth medium containing 6.5% NaCl was used for pre-enrichment and Baird Parker Agar (BPA) was used as a selective medium. Polymerase Chain Reaction technique was used to confirm the suspected colonies with the nuc gene for S. aureus and the mecA gene for MRSA. Kirby-Bauer standard disc diffusion method was applied for antibiotic susceptibility of MRSA. Results: Of the 250 investigated raw meat samples, 21.2% were positive for S. aureus, which comprised 31% beef, 14% chicken, and 16 %turkey samples. Of the 250 investigated raw meat samples, 4% were positive for MRSA, which comprised 6% beef, 3% chicken, and 2% turkey samples. All MRSA isolates were found to be resistant to penicillin, sulfamethoxazole, trimethoprim, cefoxitin, and oxacillin, but they were susceptible to vancomycin. Conclusion: In recent years, MRSA has been called a zoonotic pathogen that poses a serious risk for food safety and public health. Therefore, we believe that this study will shed light on new studies on the prevalence of MRSA in various animal-originated foods.

Occurrence and Antimicrobial Resistance of Staphylococcus aureus Isolated from Healthy Pet Rabbits.

Background: Staphylococcus aureus is a ubiquitous microorganism and an opportunistic pathogen responsible for numerous diseases in humans and animals, characterized by different clinical pictures with acute or subacute course. S. aureus, due to its great adaptability and versatility in terms of infections and hosts, can be considered a relevant pathogen because of the harmful effects on animal health and its potential for transmission from animals to humans and vice versa. In recent years, a marked increase in multidrug-resistant S. aureus has been reported, posing a serious threat for disease management, food safety, and animal and human health as they limit available therapeutic options. In light of a growing interest of the scientific community for this micro- organism and considering the limited data availability on the prevalence of this pathogen in pet rabbits, the purpose of this research was to evaluate the presence of S. aureus in pet rabbits. Materials and Methods: From November 2021 to December 2022, nasal swabs were collected from 50 pet rabbits from private households in the Campania Region, southern Italy, and underwent analysis for S. aureus detection. Samples were enriched in broth, then inoculated onto nutrient and selective media, including Blood agar base supplemented with 7% sheep blood and Baird-Parker Agar Base, following standard laboratory protocols. Incubations in aerobic conditions at 37°C were performed for 24/48h for colony identification. Antimicrobial susceptibility testing for all S. aureus isolates was conducted using the disc diffusion method. Results: Our results reported the presence of S. aureus in 16/50 (32%) rabbits examined, showing high levels of phenotypic resistance to different antibiotics, in particular penicillin 10U (81.2%) and erythromycin 15 μg (62.5%). Conclusion: The study demonstrated that pet rabbits represent a significant reservoir of S. aureus and contributes to the knowledge on the phenotypic antimicrobial resistance of these bacteria in rabbits raised in a domestic environment.

Lesões labiais em cavalos: relato de caso

ABSTRACT The article presents the results of monitoring studies on the mucous membrane lesions in Trakehner horses. Based on clinical examination, the localization, size, and number of ulcers, as well as the inflammatory reaction to the presence of necrotic tissue, were determined. In addition to clinical examination, planimetric studies were also carried out to determine the size of the ulcers and the dynamics of their reduction during treatment with aerosol application of PVP iodine spray. To determine the nature of inflammation in the lesion focus, smears were taken from the surface of the ulcers. Microbiological and mycological studies of pathological tissues and feed samples were carried out to establish the etiological factors contributing to the ulcers. It was found that the size of the ulcers varied from 1 to 6 cm, and they were located on the upper and lower lips on both the right and left sides. Neutrophilic leukocytosis was observed in the focus of inflammation. Leukocytes actively phagocytized microorganisms present in the pathological focus. Microscopic fungi of the species A. flavus, A. fumigatus, M. species, and P. commune sprouted on Saburo medium from necrotic tissues of the pathological focus. A similar composition of fungal species was obtained by sowing feed samples fed to horses. Microbiological studies of ulcers revealed the growth of two microorganisms, S. aureus and P. vulgaris, on Baird-Parker agar. On the third day of treatment, compared to the first day, the size of the ulcers decreased by 22.8 %. It is recommended to perform primary surgical treatment of the ulcer, apply PVP iodine spray once a day, and replace the affected feed with high-quality feed to achieve a positive therapeutic effect.

Ready-to-Eat Foods: A Potential Vehicle for the Spread of Coagulase-Positive Staphylococci and Antimicrobial-Resistant Staphylococcus aureus in Buea Municipality, South West Cameroon.

The consumption of ready-to-eat (RTE) foods contaminated with coagulase-positive staphylococci (CoPS) and especially Staphylococcus aureus puts consumers at a potential risk of food-borne disease or colonization and subsequent infection. This cross-sectional study determined the levels of CoPS and the presence of S. aureus in RTE foods sold in Buea municipality. A total of 420 RTE food samples, comprising 70 each of cake, bread, fruit salad, meat hot-pot, suya, and boiled rice were randomly purchased from February to August 2020. The CoPS counts were determined by culturing on Baird-Parker agar, and S. aureus was identified by amplification of the nuc gene using the polymerase chain reaction. All S. aureus isolates were screened for the presence of classical staphylococcal enterotoxin genes. To determine antimicrobial resistance profiles, each isolate was tested against 11 antimicrobials. Oxacillin-resistant S. aureus strains were analyzed for the presence of the mecA gene. Overall, 161 (38.3%) samples had detectable levels of CoPS ranging from 2.0 to 5.81 log10 CFU/g. Based on CoPS levels, 37 (8.81%) of the 420 RTE food samples-only fruit salad and meat hot-pot, had unsatisfactory microbiological quality. A total of 72 S. aureus isolates, comprising 52.78% from fruit salad, 16.67% from meat hot-pot, 12.5% from boiled rice, 9.72% from suya, 5.56% from bread, and 4.17% from cake, were recovered. None of the S. aureus isolates possessed any of the classical enterotoxin genes. All the isolates were susceptible to vancomycin and ofloxacin, while 68 (94.44%) and 66 (91.67%) were susceptible to oxacillin and ciprofloxacin, respectively. Resistance to penicillin (93.06%) was highest, followed by amoxicillin (91.67%) and erythromycin (79.17%). Four isolates were identified as methicillin-resistant S. aureus, all of which carried the mecA gene. A total of 24 antibiotypes were identified. Our findings showed that RTE foods sold in the Buea municipality are likely vehicles for the transmission of CoPS and antimicrobial-resistant S. aureus.

Pathogenic Bacterial Communities Isolated and Identified in Stingless Bee (Kelulut) Honey from Selected Farms in Terengganu

In Malaysia, stingless bees can be categorised into two genera: Melipona and Trigona, known as “kelulut”. The high demand for kelulut honey boosts the production of the honey industry. Previous studies reported that stingless bee (kelulut) honey and its products were contaminated with pathogenic bacteria during harvesting and processing. This research aims to isolate and identify the pathogenic bacteria in kelulut honey. Forty-eight samples of kelulut honey (open and closed pot) and propolis were obtained from selected farms in Terengganu by focusing on a major stingless bee species available in Malaysia, Heterotrigona itama. In addition, the swabbing technique was done on the wooden beehive of the kelulut to evaluate the environmental contamination. The pathogenic bacteria were isolated using specifically selected agar, such as Bacillus cereus agar (for B. cereus), Baird-Parker agar (for Staphylococcus aureus), and MacConkey agar (for other pathogenic bacteria), which were confirmed through a biochemical test. All samples were analysed, and the results showed that B. cereus (7/48), Pseudomonas aeruginosa (10/48), Pantoea spp. (11/48), Serratia plymuthica (6/48), and S. aureus (9/48) were obtained in the samples. This study indicates that kelulut honey was contaminated with B. cereus, P. aeruginosa, Pantoea spp., S. plymuthica, and S. aureus. Isolated pathogenic bacteria may exist in the kelulut honey through food handlers, utensils, and the environment. Hence, the stakeholders should strictly follow good standard operating procedures and guidelines by the kelulut honey industry to prevent foodborne illness.

The Correlation between Clumping Factor A Gene Expression in Biofilm Formation and Antibiotic Resistance Among Staphylococcus aureus Isolated from Urine Samples of Imam Khomeini Hospital, Tehran

Background & Objective: Staphylococcus aureus causes problems in hospitals and it has emerged as a serious agent acquired from the environment in recent years. One of the capabilities of S. aureus is the formation of biofilms, in which bacteria can exchange antibioticresistance genes among themselves and increase the virulence of other strains of this species (S. aureus). A surface protein attached to the cell wall in S. aureus clumping factor A is a virulence factor in various staphylococcal infections.
 Materials & Methods: In this study, after the Urea Analysis (UA) test, the urea culture test was applied to the blood agar and Baird-Parker Agar culture media from the infectious urine samples in Imam Hospital, Tehran, to identify S. aureus isolates. Finally, a molecular method was used for the confirmation of identified isolates. The microliter plate method was performed to determine the biofilm formation ability. The disk diffusion method was also used for profiling the antibiotic resistance of the isolates.
 Results: In the results of this study, 45 out of 160 urinary clinical samples were positive for S. aureus, among which 42 isolates expressed the clfA gene. Moreover, 39 isolates had the ability to form biofilms in vitro. Among these 42 isolates, the highest (88%) and the lowest (16%) rates of antibiotic resistance were observed against penicillin and cefoxitin, respectively. Data analysis with SPSS software and chi-square indicated a significant relationship between gene expression and biofilm production with antibiotic resistance (P < 0.05).
 Conclusion: The resistance of S. aureus bacteria is increasing strongly due to the repeated use of antibiotics such as beta-lactams, especially in respiratory infections and pharyngitis. Moreover, biofilm formation and virulence factors, such as clfA and clfB, cause concerns to the World Health Organization for treatment, especially for people with sepsis or toxemia.

Staphylococcus aureus in Horses in Nigeria: Occurrence, Antimicrobial, Methicillin and Heavy Metal Resistance and Virulence Potentials

Staphylococcus aureus was isolated from a total of 360 nasal and groin skin swabs from 180 systematic randomly-selected horses slaughtered for meat at Obollo-Afor, Enugu State, Southeast Nigeria and antimicrobial, methicillin and heavy metal resistance profile and virulence potentials of the isolates established. Baird-Parker agar with egg yolk tellurite was used for S. aureus isolation. S. aureus isolates were confirmed biochemically and serologically using a specific S. aureus Staphytect Plus™ latex agglutination test kit. The antimicrobial resistance profile, methicillin, vancomycin and inducible clindamycin resistance, and β-lactamase production of the isolates were determined with disc diffusion. Tolerance to Copper, Cadmium, Lead and Zinc was assessed using the agar dilution method and virulence potentials were determined using phenotypic methods. Forty-three (23.9%) of the 180 horses harbored S. aureus. Some 71 S. aureus were recovered from the 360 samples. Two (2.8%) of the 71 S. aureus were methicillin-resistant S. aureus (MRSA) and 69 (97.2%) were methicillin-susceptible. MRSA was recovered from 2 (1.1%) of the 180 horses. Some 9.4% of the isolates were multiple drug-resistant (MDR). The mean multiple antibiotic resistance indices (MARI) for the isolates was 0.24. Heavy metal resistance rate of the isolates ranged between 35.4-70.4%. The isolates, including the MRSA strains, displayed virulence potentials as clumping factor and catalase, gelatinase, caseinase, heamolysin, and biofilm was at the rate of 100%, 53.5%, 43.7%, 18.3% and 23.9%, respectively. This study showed that a considerable percentage of horses slaughtered in Obollo-Afor Southeastern Nigeria are potential reservoirs of virulent multiple drug- and heavy metal-resistant S. aureus, including MRSA, that could spread to humans and the environment.

Isolation and Characterization of Staphylococcus spp. and the Unintended Discovery of Non-Staphylococcal Strains Associated with Bovine Mastitis in Region IV-A, Philippines

Bovine mastitis is a common bacterial infection that causes inflammation of the mammary gland in dairy cattle. Staphylococcus is one of the most predominant and significant pathogens known to cause mastitis. Thus, this study was designed to isolate and assess the prevalence of Staphylococcus strains in dairy cattle with clinical mastitis in Region IV-A, Philippines. Baird Parker agar (BPA) supplemented with 5% egg yolk tellurite emulsion was used as selective media to differentiate coagulase-positive S. aureus and coagulase-negative staphylococcus (CNS). The isolated pathogens were phenotypically and genetically characterized for further identification. Surprisingly, out of 85 isolates, only 56.5% belong to the genus Staphylococcus and 43.5% were non-staphylococcus strains despite the use of a staphylococcus-specific selective medium. The overall most prevalent species among the isolates were S. chromogenes with an isolation rate of 21.2%. The antimicrobial resistance profiles of the isolated bacterial pathogens were also done. Several staphylococcal isolates exhibited resistance to multiple antibiotics. S. aureus showed hyper-resistance to three out of seven antibiotics – specifically streptomycin, clindamycin, and lincomycin. Surprisingly, most of the non-staphylococcal isolates exhibited hyper-resistance to multiple antibiotics. Proteus mirabilis, P. penneri, and Providencia stuartii showed almost 100% resistance and hyper-resistance to all antimicrobials tested except amoxicillin. These findings suggest that the detection of diverse species of mastitis-causing microorganisms is an important concern for the dairy sector, as different pathogens might represent different hazards and require distinct treatments, particularly on the type of antibiotics used to treat the infection. The erroneous choice of antimicrobial could unnecessarily expose the udder microbial flora to antimicrobials, thereby inadvertently promoting the emergence of multidrug-resistant pathogens that poses a serious threat to both animal and human health. Extreme antibiotic resistance detected in strains present in mastitic dairy cattle also raises a substantial public health concern, particularly when it comes to the consumption of food and food products, such as milk.

High frequency of enterotoxin encoding genes of Staphylococcus aureus isolated from food and clinical samples

BackgroundStaphylococcus aureus is recognized as an important cause of food poisoning related to the consumption of raw, undercooked, or mishandled foods worldwide.MethodsA total of 90 individual meat samples and 200 clinical specimens were collected and investigated the frequency of S. aureus and classical enterotoxin genes. The samples were cultured on Baird-Parker and Mannitol salt agar and subjected for confirmatory biochemical tests and molecular detection of femA, sea, seb, sec, sed, and see genes.ResultsA total of 31 (34.5%) meat samples and 81 (40.5%) clinical specimens were positive for the presence of S. aureus. These isolates were detected with slightly higher frequency in clinical specimens than food samples (P> 0.05). Furthermore, the frequency of S. aureus in raw meat (23.4%) was higher than that in cooked meat samples (11.1%) (P< 0.05). Staphylococcal enterotoxin (SE) genes were identified in 18 (58.1%) of 31 meat isolates and 42 (51.8%) of 81 clinical isolates. The frequency of SE genes (except see) in meat isolates was slightly higher than that in clinical isolates (P> 0.05). We found sea and see genes with higher frequency than others in both meat and clinical samples. Furthermore, 55.5% of meat isolates and 38.1% of clinical isolates possessed more than one se gene.ConclusionDetection of enterotoxigenic S. aureus in clinical and raw meat samples shows a probable risk for public health. Therefore, intensive and continuous monitoring of potentially pathogenic S. aureus is strongly recommended in order to evaluate the human health risk arising from food consumption.

Antimicrobial Resistance and Genetic Lineages of Staphylococcus aureus from Wild Rodents: First Report of mecC-Positive Methicillin-Resistant S. aureus (MRSA) in Portugal.

Simple SummaryStaphylococcus aureus is present in the microbiota of both humans and some animal species, being recognized as one of the most important opportunistic human pathogens. S. aureus is responsible for causing a variety of infections. Methicillin-resistant S. aureus (MRSA) is particularly important, as it is becoming increasingly prevalent in the population. MRSA has been increasingly reported among wild free-living animals which may impose a public health concern due to its zoonotic potential. To investigate the prevalence and antimicrobial resistance of S. aureus and MRSA in wild synanthropic rodent populations, we conducted this study on 204 rodents captured in port areas in Portugal. The antimicrobial resistance was investigated in all isolates as well as virulence genes and genetic lineages. Thirty-eight S. aureus were isolated. The results showed that six MRSA were detected with particularly interesting mecC-carrying MRSA isolates which had not yet been found in Portugal. A low frequency of antibiotic resistance and virulence genes was observed among the isolates. Nevertheless, a high diversity of clonal lineages was detected among S. aureus some of which are associated with livestock.The frequent carriage of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), by wild animals along with its zoonotic potential poses a public health problem. Furthermore, the repeated detection of the mecA gene homologue, mecC, in wildlife raises the question whether these animals may be a reservoir for mecC-MRSA. Thus, we aimed to isolate S. aureus and MRSA from wild rodents living in port areas and to characterize their antimicrobial resistance and genetic lineages. Mouth and rectal swab samples were recovered from 204 wild rodents. The samples were incubated in BHI broth with 6.5% of NaCl and after 24 h at 37 °C the inoculum was seeded onto Baird-Parker agar, Mannitol Salt agar and ORSAB (supplemented with 2 mg/L of oxacillin) plates. Species identification was confirmed by MALDI-TOF MS. The antimicrobial susceptibility testing was performed by the Kirby–Bauer disc diffusion method against 14 antibiotics. The presence of virulence and resistance genes was performed by PCR. The immune evasion cluster (IEC) system was investigated in all S. aureus. All isolates were characterized by MLST, spa- and agr typing. From 204 samples, 38 S. aureus were isolated of which six MRSA were detected. Among the six MRSA isolates, three harbored the mecC gene and the other three, the mecA gene. All mecC-MRSA isolates were ascribed to sequence type (ST) 1945 (which belongs to CC130) and spa-type t1535 whereas the mecA isolates belonged to ST22 and ST36 and spa-types t747 and t018. Twenty-five S. aureus were susceptible to all antibiotics tested. S. aureus isolates were ascribed to 11 MLST and 12 spa-types. S. aureus presents a great diversity of genetic lineages in wild rodents. This is the first report of mecC-MRSA in Portugal.

Molecular detection of Staphylococcus aureus enterotoxin genes isolated from mastitic milk and humans in El-Behira, Egypt

Background and Aim: Milk is a chief source of many nutrients. However, we must also bear in mind that it is a potential source for many cases of food poisoning. This study was conducted to investigate the prevalence of cow mastitis and evaluate the presence of enterotoxins and antibiotic resistance patterns in Staphylococcus aureus isolated from milk and contact humans in El-Behira Province, Egypt. Materials and Methods: A total of 680 milk samples from 170 cows and 86 human samples consisting of 43 hand swabs and 43 nasal swabs were analyzed. The milk samples were subjected to the California mastitis test. Results: The general occurrence was 23.1% (157/680) where 48 quarters had clinical mastitis and 109 had subclinical mastitis. Subsequently, S. aureus was isolated in Baird-Parker agar where typical and atypical colonies were selected and submitted to coagulase and complementary tests. Out of 48 samples of mastitic milk studied, 16 (33.3%) showed contamination by S. aureus whereas 109 samples of subclinical mastitis showed contamination in only 18 (16.5%). On the opposite hand, of the 86 human samples, 33 revealed S. aureus contamination, corresponding to 38.37% of the samples. Furthermore, multiplex polymerase chain reaction targeting nuc and the staphylococcal enterotoxin-encoding genes sea, seb, sec, sed, and see were performed after culture, revealing that 88.2% (30/34) of milk samples and 93.9% (31/33) of human samples were variably positive to those genes. Conclusion: The use of nuc gene based PCR is an accurate and rapid method for S aureus isolates detection. A high prevalence of multiple drug-resistant isolates of S. aureus recovered from both human and milk represents further evidence for possible veterinary hazards as well as public hazards, especially to those that consume milk from this region.

Effect of gluten on the quality of meat products

Article Details: Received: 2020-10-14 | Accepted: 2020-11-27 | Available online: 2021-01-31 https://doi.org/10.15414/afz.2021.24.mi-prap.80-84 The aim of the study was the evaluation of gluten quality of selected meat products. Gluten is a vegetable protein, which, due to its properties, is very often used in various sectors of the food industry and therefore in the production of meat products. Sensory evaluation, microbiological examination and gluten detection were performed in samples of sausages, Poultry frankfurter, Fine salami and Malokarpatská salami. The meat products came from different producers and were purchased from a retail network. Sensory quality was determined by assessing color, aroma, juiciness, fragility and taste using a 5-point rating scale. The microbiological examination included the determination of the total number of microorganisms, the number of psychrotrophic, coliform microorganisms and coagulase-positive staphylococci. The presence of gluten alone was detected using a commercially produced GlutenTox Home test kit as well as a PCR reaction. Individual testing of selected meat products showed minor differences in the overall quality of gluten free and gluten-free meat products. Keywords: meat products, salami, gluten, quality, microbiological examination References Abaffyová, Z. et al. (2015). Cereal grain a little different. Bratislava. Pediatria pre prax . 4, 140–146. Frič, P., Keil, R. (2011). Celiac disease for practice. Olomouc. Medicína pro praxi. 8, 354–359. Hulín, P., Dostálek, P., Hochel, I. (2008). Methods for determination of gluten proteins in food. Metódy stanovení lepkových bílkovin v potravinách. Chemické listy ,102(5), 327–337. https://www.academia.edu/18759054/Methods_for_determination_of_gluten_proteins_in_food Olexová, L. et al. (2006). Detection of gluten-containing cereals in flours and ‘‘gluten-free’’ bakery products by polymerase chain reaction. Food Control , 17(3), 234–237. https://doi.org/10.1016/j.foodcont.2004.10.009 STN EN ISO 4833-1. Microbiology of food chain. Horizontal method for the enumeration of microorganisms. Part 1: Colony count at 30 degrees C by the pour plate technique. (ISO 4833-1: 2013) STN EN ISO 6888-1. Microbiology of food and animal feeding stuffs. Horizontal method for the enumeration of coagulase-positive staphylococci ( Staphylococcus aureus and other species). Part 1: Technique using Baird-Parker agar medium (ISO 6888-1:1999) STN EN ISO 4832. Microbiology. General guidance for the enumeration of coliforms. Colony count technique Taylor, S. (1992). Chemistry and detection of food allergens. Food Technology , 46(5), 148–152. https://doi.org/10.1080/10408399609527761 Toldrá, F. (2002). Fermentation and starter cultures. Dry-cured meat products , Trumbull : Food &amp; Nutrition Press Inc., 89–112. https://doi.org/10.1002/9780470385111 van Hengel, A. J. (2007). Food allergen detection methods and the challenge to protect food-allergic consumers. Analytical and Bioanalytical Chemistry , 389(1), 111–118. https://doi.org/10.1007/s00216-007-1353-5

IDENTIFICATION AND OCCURRENCE OF ANTIBIOTIC RESISTANCE OF Staphylococcus aureus AND Escherichia coli ISOLATED FROM RECREATIONAL PARKS AROUND KOTA KINABALU, SABAH

Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) are contributors to infection cases among the Asian population. S. aureus is found in the mucous lining of noses and is mainly non-pathogenic while E. coli, mostly harmless bacteria, are found in the intestine. Pathogenic strains of both bacteria have adverse effects on the elderly and younger age group of the population. Samples were collected from recreational parks around Kota Kinabalu as they are hotspots frequently visited by families with both age groups. The bacterial samples were isolated and cultured on selective media such as Baird-Parker agar (BPA), Brain Heart Infusion (BHI) agar, MacConkey agar and Eosin-Methylene Blue (EMB) agar. Morphological characteristics of bacterial growth were observed, where S. aureus had black-shiny growth in BPAand E. coli had a metallic-green sheen in EMB agar. The suspected bacteria samples were then stained and viewed under a light microscope. S. aureus was identified as gram-positive, stained violet with a circular shape and clustered appearance. E. coli was identified as gram-negative, stained red, rod-shaped with 2 – 3 bacterial alignment. Antibiotic resistance test resulted in S. aureus and E. coli samples did not display 100% resistance among 4 antibiotics tested (ampicillin, penicillin, tetracycline and chloramphenicol). Most of the bacteria samples were a minimum inhibitory of 0.1 mg/mL of antibiotic concentration. These results provide a foundation for further research on identifying bacterial strains using molecular methods. The findings can then be used to disseminate information to the public to create awareness of potential disease outbreaks in the city.

Chemical and Microbiological Properties of Kefir Produced by Kefir Grains in Raw and Pasteurized Cow’s Milk

The traditional production of kefir uses raw milk as a fermentation substrate which is not acceptable due to food safety issues. However, kefir grains were reported to have antimicrobial properties. Hence, this study determined if kefir grains inhibited certain spoilage and pathogenic microorganisms by comparing the chemical and microbiological properties of the initial substrates: raw milk (RM) and pasteurized milk (PM) to the final products: raw milk kefir (RMK) and pasteurized milk kefir (PMK). Both final products had a significant decrease in fat, moisture, and pH, and a significant increase in protein, total solids, and titratable acidity than RM and PM, respectively. Total solids, titratable acidity, pH, total alcohol, and ethanol were significantly higher in PMK than RMK. RMK had a significantly lower coliform count than RM while both RMK and PMK had significantly higher lactic acid bacteria, yeast and mold count than RM and PM, respectively. Kefir grains’ microbial inhibiting activity was examined if plates were positive or negative against spoilage and pathogenic microorganisms. Escherichia coli-positive plates in Levine’s eosin methylene blue agar decreased in RMK. Salmonella spp.-positive plates in bismuth sulphite agar (BSA), Hektoen enteric agar, and xylose lysine deoxycholate agar decreased in RMK and PMK except for RMK in BSA. Staphylococcus aureus-positive plates in Baird-Parker agar decreased in RMK. RMK and PMK attained the kefir’s standard values for the chemical composition, lactic acid bacteria, yeast and mold count. Nonetheless, the mere presence of spoilage and pathogenic microorganisms in the final products made it unsafe for consumption.

Inhibition of Staphylococcus aureus LC554891 by Moringa oleifera Seed Extract either Singly or in Combination with Antibiotics.

Bacterial outbreaks caused by Staphylococcus aureus (S. aureus) are interesting due to the existence of multidrug resistant (MDR) isolates. Therefore, there is a need to develop novel ways to control such MDR S. aureus. In this study, some natural agents such as honey bee (HB), extracts of either Moringa oleifera seeds (MSE), or leaves (MLE) and essential oils of garlic, clove, and moringa were studied for their inhibitory activity against this S. aureus pathogen. About 100 food samples including beef luncheon (n = 25), potato chips (n = 50), and corn flakes (n = 25) were investigated for possible pollution with the S. aureus bacteria. The isolated bacteria suspected to belong S. aureus that grew well onto Baird–Parker agar (Oxoid) and shiny halo zones and positive coagulase reaction were selected and identified by API-Kits; all of them that were approved belong to S. aureus (18 strains). The sensitivity of the obtained 18 S. aureus bacterial strains to 12 antibiotics were evaluated; all of them were resistant to ofloxacin; however, other antibiotics tested showed variable results. Interestingly, the S. aureus No. B3 isolated from beef luncheon was resistant to 10 antibiotics out of 12 ones tested. Multiple antibiotic resistance index (MAR) of this S. aureus strain was about 83.3%. Therefore, its identification was confirmed by sequencing of a 16S rRNA gene which approved a successful biochemical identification carried out by API Kits and such strain was designated S. aureus LC 554891. The genome of such strain appeared to contain mecA gene encoding methicillin resistance; it was found to contain hla, hlb, tsst-1, and finbA that encode α-blood hemolysis, β-blood hemolysis, toxic shock syndrome gene, and fibrinogen-binding protein gene, respectively. In addition, the virulence factors viz. sea; seb; sec encoding enterotoxins were detected in the DNA extracted from S. aureus B3 strain. Aqueous extract of Moringa oleifera seeds (MSE) showed inhibitory activity against S. aureus LC 554891 better than that obtained by tetracycline, essential oils or HB. Minimum inhibitory concentration (MIC) of MSE was 20µg/mL. Instrumental analysis of MSE showed 14 bioactive chemical compounds. Combinations of both MSE and tetracycline showed distinctive inhibitory activity against S. aureus LC 554891 than that obtained by either tetracycline or MSE singly.

Evaluating the Behavior of Staphylococcus aureus and Bacillus cereus in Dairy- and Non-Dairy–Based Aqueous Slurries during Manufacturing of Table Spreads

High-moisture slurries used in the production of table spreads may permit growth of Staphylococcus aureus and Bacillus cereus and subsequent production of heat-stable enterotoxins. Compliance with the Food Safety Modernization Act (FSMA), specifically 21 CFR Part 117, subpart B and section 117.80 (c)(2) and (c)(3), requires a hazard analysis to determine whether preventive controls are needed. This study estimates the risk of potential growth of S. aureus and B. cereus in eight different dairy- and non-dairy-based slurries during extended storage and use. Mathematical models were used to screen which slurries might support the growth of S. aureus and B. cereus. Samples were individually inoculated with multiple strains of S. aureus and B. cereus to achieve a target level of 102 to 103 CFU/g. Inoculated and uninoculated slurry samples were incubated at typical holding temperatures of 35°C (95°F), 46.1°C (115°F), and 54.4°C (130°F). Samples were removed and tested following inoculation (time zero), after 4 and 12 h, and after 1, 2, 3, 4, 5, 6, 7, and 10 days of incubation at the target temperatures. All experiments were repeated in triplicate. Samples were analyzed for S. aureus and B. cereus using Baird-Parker agar and mannitol yolk polymyxin agar, respectively. Neither S. aureus nor B. cereus exceeded (P < 0.05) proposed food safety limits (105 CFU/g) at the evaluated experimental conditions. The study highlights the role of multiple hurdles (e.g., pH, potassium sorbate and sodium benzoate, salt, and other ingredients) in assuring microbiological safety of in-process dairy- and non-dairy-based slurries used in the production of table spreads. This study also found that mathematical models representative of product composition, intrinsic parameters, and experimental conditions can help risk managers make informed decisions during product development. Finally, the study findings indicate no significant risk of growth of the target pathogens associated with the dairy- and non-dairy-based aqueous slurries used in the routine manufacturing of table spreads.

Süt kaynaklı Staphylococcus aureus suşları gelişimi üzerine peynirdeki nisinin etkisi

The use of bacteriocins in protecting the foods has significantly increased in recent years. Nisin is an important bacteriocin and is a food preservative additive safely used in food industry. In the present study, it was aimed to investigate the effect of nisin, which is an alternative to the synthetic additives, on the growth of milk-derived Staphylococcus aureus strains in the cheese. For this purpose, 100 different milk specimens were obtained, transferred using cold chain (at 0-4°C), and inoculated to Baird-Parker agar. At the end of inoculation process, 37 suspicious colonies were selected and coagulase-positive Staphylococcus aureus was detected. The strains obtained were passaged to the pre-enriched agars and the intensities of the strains were determined according to McFarland. Besides these strains, the coagulase-positive Staphylococcus aureus strains obtained from Etlik Veterinary Microbiology Institute were inoculated into each specimen at 104 CFU mL-1 concentration. The cheese production was made using milks containing the strains, which were isolated from the milk, and the standard strains at different concentrations. The specimens obtained from cheeses produced from milk containing using standard strains were divided into groups K, A, B, and C containing 0, 100, 200, and 400 IU mL-1 nisin, respectively. Similarly, the specimens obtained from cheeses produced from milks containing milk-derived strains were divided into groups K1, A1, B1, and C1 containing 0, 100, 200, and 400 IU mL-1 nisin, respectively. The procedures were triplicated. Staphylococcus aureus counting on the cheese specimens were performed on 1st, 6th, 12th, and 24th hours from curd to ball and on 5th, 10th, 20th, 30th, and 40th days of storage at +4°C. In conclusion, it was observed that the nisin inhibited the growth of all the Staphylococcus aureus strains. However, it was also determined that the inhibition of milk-derived Staphylococcus aureus strains were higher than the inhibition of standard strains.

Antibiotic Resistance and Biofilm Production in Catalase-Positive Gram-Positive Cocci Isolated from Brazilian Pasteurized Milk

Background: Milk is a reservoir for several groups of microorganisms, which may pose health risks. The aim of this work was to assess the antibiotic resistance and biofilm production in catalase-positive Gram-positive cocci isolated from Brazilian pasteurized milk. Methods: The bacteria were isolated using Baird-Parker agar and identified by MatrixAssisted Laser Desorption/Ionization-Time-Of-Flight (MALDI-TOF) mass spectrometer. Disk diffusion technique was used to evaluate antimicrobial susceptibility. For qualitative evaluation of biofilm production, the growth technique was used on Congo Red Agar. Results: Totally, 33 out of 64 isolates were identified, including Staphylococcus epidermidis (n=3; 4.7%), Macrococcus caseolyticus (n=14; 21.9%), and Kocuria varians (n=16; 25%). Twenty-two isolates were resistant to at least one antibiotic. Biofilm production was detected in only 5 isolates of K. varians and 1 isolate of S. epidermidis. All 14 M. caseolyticus isolates were resistant to at least one antibiotic; but, multidrug resistant (MDR) isolates were not detected. Among all K. varians isolates, 4 were resistant to at least one antibiotic from three different classes and were considered to be MDR. Conclusion: The presence of antibiotic-resistant M. caseolyticus, S. epidermidis, and K. varians isolates, especially MDRs, in milk samples highlights the possible role of milk as a reservoir of resistance genes