Search results for: isolates

Authors: Ahlem Nefzi, Rania Aydi Ben Abdallah, Hayfa Jabnoun-Khiareddine, Ammar Nawaim, Rabiaa Haouala, Mejda Daami-Remadi

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Seven endophytic fungi were isolated from the wild Solanaceous species Lycium arabicum growing in the Tunisian Centre-East and were assessed for their ability to suppress Fusarium Crown and Root Rot disease caused by Fusarium oxysporum f. sp. radicis lycopersici (FORL) and to enhance plant growth. Fungal isolates were shown able to colonize tomato cv. Rio Grande roots, crowns, and stems. A significant promotion in all studied growth parameters (root length, shoot height, and roots and shoots fresh weight) was recorded in tomato plants treated with fungal conidial suspensions or their cell-free culture filtrates compared to FORL-inoculated or pathogen-free controls. I15 and I18 isolates were shown to be the most effective leading to 85.7-87.5 and 93.6-98.4% decrease in leaf and root damage index and the vascular discoloration extent, respectively, over FORL-inoculated and untreated control. These two bioactive and growth-promoting isolates (I15 and I18) were morphologically characterized and identified using rDNA sequencing gene as being Alternaria alternata (MF693801) and Fusarium fujikuroi (MF693802). These fungi significantly suppressed FORL mycelial growth and showed chitinolytic, proteolytic and amylase activities whereas only F. fujikuroi displayed a lipolytic activity. This study clearly demonstrated the potential use of fungi naturally associated with L. arabicum as biocontrol and bio-fertilizing agents.

Keywords: biocontrol, endophytic fungi, Fusarium oxysporum f. sp. radicis-lycopersici, tomato promotion, Lycium arabicum

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Authors: Sima E. Rugarabamu, Mecky I. Matee, Elison N. M. Simon

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Background: In Tanzania bacteriological studies of etiological agents of oro-facial infections are very limited, and very few have investigated anaerobes. The aim of this study was to determine the spectrum of bacterial agents involved in oral and maxillofacial infections in patients attending Muhimbili National Hospital, Dar-es-salaam, Tanzania. Method: This was a hospital based descriptive cross-sectional study that was conducted in the Department of Oral and Maxillofacial Surgery of the Muhimbili National Hospital in Dar es Salaam, Tanzania from 1st January 2014 to 31st August 2014. Seventy (70) patients with various forms of oral and maxillofacial infections who were recruited for the study. The study participants were interviewed using a prepared questionnaire after getting their consent. Pus aspirate was cultured on Blood agar, Chocolate Agar, MacConkey agar and incubated aerobically at 37°C. Imported blood agar was used for anaerobic culture whereby they were incubated at 37°Cin anaerobic jars in an atmosphere of generated using commercial gas-generating kits in accordance with manufacturer’s instructions. Plates were incubated at 37°C for 24 hours (For aerobic culture and 48 hours for anaerobic cultures). Gram negative rods were identified using API 20E while all other isolates were identified by conventional biochemical tests. Antibiotic sensitivity testing for isolated aerobic and anaerobic bacteria was detected by the disk diffusion, agar dilution and E-test using routine and commercially available antibiotics used to treat oral facial infections. Results: This study comprised of 41 (58.5%) males and 29 (41.5%) females with a mean age of 32 years SD +/-15.1 and a range of 19 to 70 years. A total of 161 bacteria strains were isolated from specimens obtained from 70 patients which were an average of 2.3 isolates per patient. Of these 103 were aerobic organism and 58 were strict anaerobes. A complex mix of strict anaerobes and facultative anaerobes accounted for 87% of all infections.The most frequent aerobes isolated was streptococcus spp 70 (70%) followed by Staphylococcus spp 18 (18%). Other organisms such as Klebsiella spp 4 (4%), Proteus spp 5 (5%) and Pseudomonas spp 2 (2%) were also seen. The anaerobic group was dominated by Prevotella spp 25 (43%) followed by Peptostreptococcus spp 18 (31%); other isolates were Pseudomonas spp 2 (1%), black pigmented Pophyromonas spp 4 (5%), Fusobacterium spp 3 (3%) and Bacteroides spp 5 (8%). Majority of these organisms were sensitive to Amoxicillin (98%), Gentamycin (89%), and Ciprofloxacin (100%). A 40% resistance to metronidazole was observed in Bacteroides spp otherwise this drug and others displayed good activity against anaerobes. Conclusions: Oral and maxillofacial facial infections at Muhimbili National Hospital are mostly caused by streptococcus spp and Prevotella spp. Strict anaerobes accounted for 36% of all isolates. The profile of isolates should assist in selecting empiric therapy for infections of the oral and maxillofacial region. Inclusion of antimicrobial agents against anaerobic bacteria is highly recommended.

Keywords: bacteria, oral and maxillofacial infections, antibiotic susceptibility, Tanzania

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Authors: Seyed Reza Aghili, Tahereh Shokohi, Shirin Sadat Hashemi Fesharaki, Mohammad Ali Boroumand, Bahar Salmanian

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Introduction: Intravenous catheter-associated fungal infection as nosocomial infection continue to be a deep problem among hospitalized patients, decreasing quality of life and adding healthcare costs. The capacity of catheters in the spread of candidemia in heart failure patients is obvious. The aim of this study was to evaluate the prevalence and genetic identification of Candida species in heart disorder patients. Material and Methods: This study was conducted in Tehran Hospital of Cardiology Center (Tehran, Iran, 2014) during 1.5 years on the patients hospitalized for at least 7 days and who had central or peripheral vein catheter. Culture of catheters, blood and skin of the location of catheter insertion were applied for detecting Candida colonies in 223 patients. Identification of Candida species was made on the basis of a combination of various phenotypic methods and confirmed by sequencing the ITS1-5.8S-ITS2 region amplified from the genomic DNA using PCR and the NCBI BLAST. Results: Of the 223 patients samples tested, we identified totally 15 Candida isolates obtained from 9 (4.04%) catheter cultures, 3 (1.35%) blood cultures and 2 (0.90%) skin cultures of the catheter insertion areas. On the base of ITS region sequencing, out of nine Candida isolates from catheter, 5(55.6%) C. albicans, 2(22.2%) C. glabrata, 1(11.1%) C. membranifiaciens and 1 (11.1%) C. tropicalis were identified. Among three Candida isolates from blood culture, C. tropicalis, C. carpophila and C. membranifiaciens were identified. Non-candida yeast isolated from one blood culture was Cryptococcus albidus. One case of C. glabrata and one case of Candida albicans were isolated from skin culture of the catheter insertion areas in patients with positive catheter culture. In these patients, ITS region of rDNA sequence showed a similarity between Candida isolated from the skin and catheter. However, the blood samples of these patients were negative for fungal growth. We report two cases of catheter-related candidemia caused by C. membranifiaciens and C. tropicalis on the base of genetic similarity of species isolated from blood and catheter which were treated successfully with intravenous fluconazole and catheter removal. In phenotypic identification methods, we could only identify C. albicans and C. tropicalis and other yeast isolates were diagnosed as Candida sp. Discussion: Although more than 200 species of Candida have been identified, only a few cause diseases in humans. There is some evidence that non-albicans infections are increasing. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition are considered to be associated with candidemia in hospitalized heart failure patients. Identifying the route of infection in candidemia is difficult. Non-albicans candida as the cause of candidemia is increasing dramatically. By using conventional method, many non-albicans isolates remain unidentified. So, using more sensitive and specific molecular genetic sequencing to clarify the aspects of epidemiology of the unknown candida species infections is essential. The positive blood and catheter cultures for candida isolates and high percentage of similarity of their ITS region of rDNA sequence in these two patients confirmed the diagnosis of intravenous catheter-associated candidemia.

Keywords: catheter-associated infections, heart failure patient, molecular genetic sequencing, ITS region of rDNA, Candidemia

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Authors: Omaima A. Sharaf, Tarek A. Moussa, Said M. Badr El-Din, H. Moawad

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Background: Polycyclic aromatic hydrocarbons (PAHs) pose great environmental and human health concerns for their widespread occurrence, persistence, and carcinogenic properties. PAHs releases due to anthropogenic activities to the wider environment have led to higher concentrations of these contaminants than would be expected from natural processes alone. This may result in a wide range of environmental problems that can accumulate in agricultural ecosystems, which threatened to become a negative impact on sustainable agricultural development. Thus, this study aimed to evaluate the physico-chemical, and microbial properties of the compost leachate (CL) to assess its role as nutrient and microbial source (biostimulation/bioaugmentation) for developing a cost-effective bioremediation technology for PAHs contaminated sites. Material and Methods: PAHs-degrading bacteria were isolated from CL that was collected from a composting site located in central Scotland, UK. Isolation was carried out by enrichment using phenanthrene (PHR), pyrene (PYR) and benzo(a)pyrene (BaP) as the sole source of carbon and energy. The isolates were characterized using a variety of phenotypic and molecular properties. Six different isolates were identified based on the difference in morphological and biochemical tests. The efficiency of these isolates in PAHs utilization was assessed. Further analysis was performed to define taxonomical status and phylogenic relation between the most potent PAHs-utilizing bacterial strains and other standard strains, using molecular approach by partial 16S rDNA gene sequence analysis. Results indicated that the 16S rDNA sequence analysis confirmed the results of biochemical identification, as both of biochemical and molecular identification of the isolates assigned them to Bacillus licheniformis, Pseudomonas aeruginosa, Alcaligenes faecalis, Serratia marcescens, Enterobacter cloacae and Providenicia which were identified as the prominent PAHs-utilizers isolated from CL. Conclusion: This study indicates that the CL samples contain a diverse population of PAHs-degrading bacteria and the use of CL may have a potential for bioremediation of PAHs contaminated sites.

Keywords: polycyclic aromatic hydrocarbons, physico-chemical analyses, compost leachate, microbial and biochemical analyses, phylogenic relations, 16S rDNA sequence analysis

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Authors: D. S. V. Barbieri, R. R. Gomes, G. D. Santos, P. F. Herkert, M. Moreira, E. S. Trindade, V. A. Vicente

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The importance of yeast infections has increased in recent decades. The monitoring of Candida yeasts has been relevant in the study of groups and populations. This research evaluated 31 Candida spp. isolates from oral microbiota of 12 Brazilian schoolchildren coinfected with Streptococcus mutans. The isolates were evaluated for their ability to form biofilm in vitro and molecularly characterized based on the sequencing of intergenic spacer regions ITS1-5,8S-ITS2 and variable domains of the large subunit (D1/D2) regions of the rDNA, as well as ABC system genotyping. The sequencing confirmed 26 lineages of Candida albicans, three Candida tropicalis, one Candida guillhermondii and one Candida glabrata. Genetic variability and differences on in biofilm formation were observed among Candida yeasts lineages. At least one Candida strain from each caries activity child was C.albicans genotype A or Candida non-albicans. C. tropicalis was associated with highest cavities rates. These results indicate that the presence of C. albicans genotype A or multi-colonization by non albicans species seem to be associates to the potentialization of caries risk.

Keywords: biofilm, Candida albicans, oral microbiota, caries

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Authors: Latifat Afolake Ogunfolabo, Hakeem Babafemi Ogunfolabo

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The gastrointestinal tract is densely populated with micro-organism which closely and intensively interacts with the host and ingested feed. Food borne infections are some of the major international challenges that lead to high mortality and also, antimicrobial resistance, which has been classified as a serious threat by World Health Organization. Samples of slaughtered cattle and goats intestines were collected and standard culture methods were used for bacteria isolation and identification. Minimum inhibitory concentration of commonly used antibiotic using modification of the disk diffusion method was carried out on isolates. The samples cultured were all positive to Pseudomonas aeruginosa (95% and 90%), Escherichia coli (85%), Salmonella typhi (70% and 60%), Staphylococcus aureus (75%and 100%), Micrococcus luteus (55% and35%), Bacillus macerans (60% and 5%), Bacillus cereus (25% and 20%), Clostridium perfringens (20% and 5%), Micrococcus varians (20% and 5%), Bacillus subtilis (25% and 5%), Streptococcus faecalis (40% and 25%) and Streptococcus faecium (15% and 10%) in goat and cattle respectively. Also, Proteus mirabilis (40%), Micrococcus luteus (35%), Proteus vulgaris (30%), Klebsiella aerogenes(15%) were isolated from cattle. The total coliform (13.55 x10⁵cfu/gm ± 1.77) and (20.30 x10⁵cfu/gm ± 1.27) counts were significantly higher than the total bacteria count (8.3 x10⁵cfu/gm ± 1.41) and (16.60 x10⁵cfu/gm ±0.49) for goat and cattle respectively. Selected Bacteria count of isolates showed that Staphylococcus aureus had the highest significant value (6.9 x10⁵cfu/gm ± 0.57) and (16.80 x10⁵cfu/gm ± 0.57) Escherichia coli (4.60 x10⁵cfu/gm ± 0.42) and (7.05 x10⁵cfu/gm ± 0.64) while the lowest significant value was obtained in Salmonella/Shigella (1.7 x10⁵cfu/gm ± 0.00) and (1.5 x10⁵cfu/gm ± 0.00) for goat and cattle respectively. Susceptibility of bacteria isolated from slaughtered goat and cattle intestine to commonly used antibiotics showed that the highest statistical significant value for zone of inhibition for goat was obtained for Ciprofloxacin (30.00 ± 2.25, 23.75 ± 2.49, 17.17 ± 1.40) followed by Augmentin (28.33 ± 1.22, 21. 83 ± 2.44, 16.67 ± 1.49), Erythromycin (27.75 ±1.48, 20.25 ± 1.29, 16.67 ± 1.26) while the lowest values were obtained for Ofloxacin (27.17 ± 1.89, 21.42 ± 2.19, 16.83 ± 1.26) respectively and values obtained for cattle are Ciprofloxacin (30.64 ± 1.6, 25.79 ± 1.76, 8.07 ± 11.49) followed by Augmentin (28.29 ± 1.33, 22.64 ± 1.82, 17.43 ± 1.55) Ofloxacin (26.57 ± 2.02, 20.79 ± 2.75, 16.21 ± 1.19) while the lowest values were obtained for Erythromycin (26.64 ± 1.49, 20.29 ± 1.49, 16.29 ± 1.33) at different dilution factor (10⁻¹, 10⁻², 10⁻³) respectively. The isolates from goat and cattle were all susceptible to Augmentin at the three different dilution factors. Some goat isolates are intermediate to Ciprofloxacin and Erythromycin at 10⁻² and 10⁻³, while resistance to Ciprofloxacin at 10⁻³ dilution factor. Ciprofloxacin and Ofloxacin at the dilution factors of 10⁻³ and 10⁻¹ for some cattle isolate and resistance were observed for Ofloxacin and Erythromycin at dilution of 10⁻³. These results indicate the susceptibilities and the antimicrobial resistance to commonly used antibiotic.

Keywords: antibiotic susceptibility, bacteria, cattle, goat, identification

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Authors: Meenakshi Chaudhary, V .S. Randhawa, M. Jais, R. Dutta

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Introduction: An outbreak of Multi drug resistant S. Typhi (i.e. resistance to chloramphenicol, ampicillin, and trimethoprim-sulfamethoxazole) occurred in 1990's in India which peaked in 1992-93 and resulted in the change of drug of choice from chloramphenicol to ciprofloxacin for enteric fever. Currently an emergence of Ciprofloxacin susceptible S. Typhi isolates in the region is being reported which appears to be chromosomally mediated. Methodology: Six hundred sixty four strains were randomly selected from the time period between January 2008-December 2011 at the National Salmonella Phage Typing Centre, LHMC, New Delhi. The strains were representative of the north, central and south zones of India. All isolates were subjected to serotyping, biotyping, phage typing and then to antimicrobial susceptibility testing by CLSI disk diffusion (CLSI) technique to Ciprofloxacin, Cefotaxime, Ampicillin, Chloramphenicol, Trimethoprim-Sulfomethoxazole and Tetracycline. Subsequently MIC of the isolates was determined by E-test (AB-Biodisc). Results: More than 80% of the tested strains had intermediate susceptibility to ciprofloxacin. The E test revealed the MIC (Ciprofloxacin) of these strains to be in the range of 0.12 to 0.5 µg/ml. Sixty nine percent of ciprofloxacin intermediate susceptible strains belonged to Phage type E1 and fourteen percent of these were Vi- Negative i.e these could not be typed by the phage typing scheme of Craigie and Yen. All the strains remained susceptible to cefotaxime. Conclusion: Predominant isolation of intermediate susceptible S. Typhi strains from India would alter the recommendations of empiric treatment of enteric fever in the region. Alternative to the low cost ciprofloxacin will have to be sought or increased dosage and/or duration of ciprofloxacin will have to be recommended. The reasons for the trend of increase in percentage of intermediate susceptible S. Typhi strains are not clear but may be attributed partly to the revision of CLSI guidelines in 2013.

Keywords: salmonella typhi, decreased ciprofloxacin susceptibility, ciprofloxacin, minimum inhibitory concentration

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Authors: Moses Ikechukwu Benjamin

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The continued increase in methicillin-resistant Staphylococcuspseudintermedius (MRSP) among dogs and the zoonotic transmission event of MRSP from dogs to humans threaten veterinary medicine and public health. The cardinal objective of this study was to determine the antibiogram and frequency of toxingenes in MRSP obtained from shelter dogs with skin infections and dog owners in Abakaliki, Eastern Nigeria. Skinswabs from 61 shelter dogs with skin infections and 33 nasal swabs from dog owners were processed and analyzed using standard microbiological techniques. Susceptibility to antibiotics was determined by Kirby Bauer disc diffusion technique. The screening for Seccanine, lukD, siet, and exitoxin genes was carried out by PCR. A total of 23 (37.7 %) and 1 (3 %) MRSP strains were obtained from shelter dogs and dog owners, respectively. Generally, isolates exhibited high resistance to amoxicillin-clavulanic acid, ceftazidime, and cefepime (100 % - 66.7 %) but were very susceptible (100 % - 70.7 %) to chloramphenicol and doripenem. The only isolate from dog owners harbouredseccanine, lukD, and siet toxin genes while solatesfrom shelter dogs harbouredseccanine16 (69.6 %), lukD 17 (73.9 %), siet 20 (87 %), and exi1 (4.4 %) toxin genes. Isolates were generally observed to be more resistant than other reports from the literature. Interesting, there was a similarity in the resistance antibiotypes and frequency of toxin genes harboured by MRSP isolates between shelter dogs with skin infections and their owner in a sampled household, thus suggesting a likely zoonotic transmission event. This report of the occurrence of MRSP and high frequency of toxin genes (Seccanine,lukD, and siet) in shelter dogs and dog owners represent a major challenge, especially in terms of antibiotic therapy, and is a serious concern for both animal and public health.

Keywords: methicillin-resistant S. pseudintermedius, zoonotic transmission, antibiotic resistance, companion dogs, toxin genes

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Authors: Yazeed Al-Sheikh

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Pregnancy represents a risk factor in the occurrence of vulvovaginal candidiasis. To investigate the prevalence rate of vaginal carriage of Candida species in Saudi pregnant and non-pregnant women, high vaginal swab (HVS) specimens (707) were examined by direct microscopy (10% KOH and Giemsa staining) and parallel cultured on Sabouraud Dextrose Agar (SDA) as well as on “CHROM agar Candida” medium. As expected, Candida-positive cultures were frequently observed in pregnant-test group (24%) than in non-pregnant group (17%). The frequency of culture positive was correlated to pregnancy (P=0.047), parity (P=0.001), use of contraceptive (P=0.146), or antibiotics (P=0.128), and diabetic-patients (P < 0.0001). Out of 707 HVS examined specimens, 157 specimens were yeast-positive culture (22%) on Sabouraud Dextrose Agar or “CHROM agar Candida”. In comparison, the sensitivities of the direct 10% KOH and the Giemsa stain microscopic examination methods were 84% (132/157) and 95% (149/157) respectively but both with 100% specificity. As for the identity of recovered 157 yeast isolates, based on API 20C biotype carbohydrate assimilation, germ tube and chlamydospore formation, C. albicansand C. glabrata constitute 80.3 and 12.7% respectively. Rates of C. tropicalis, C. kefyr, C. famata or C. utilis were 2.6, 1.3, and 0.6% respectively. Sachromyces cerevisiae and Rhodotorula mucilaginosa yeasts were also encountered at a frequency of 1.3 and 0.6% respectively. Finally, among all recovered 157 yeast-isolates, strains resistant to ketoconazole were not detected, whereas 5% of the C. albicans and as high as 55% of the non-albicans yeast isolates (majority C. glabrata) showed resistance to fluconazole. Our findings may prove helpful for continuous determination of the existing vaginal candidiasis causative species during pregnancy, its lab-diagnosis and/or control and possible measures to minimize the incidence of the disease-associated pre-term delivery.

Keywords: vaginal candidiasis, Candida spp., pregnancy, risk factors, API 20C-yeast biotypes, giemsa stain, antifungal agents

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Authors: Nwafia Ifeyinwa Nkeiruka, Nwafia Walter Chukwuma

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Since the early 1980’s fungi have emerged as major causes of human diseases, especially among immunocompromised. The most commonly isolated yeast is Candida albicans and constitutes the 4th most common nosocomial BSI in humans. It is progressive and cumulative and become more complex over time.It can even lead to leaky gut syndrome that causes food and environmental allergies. It is worthy of note that all the available data on oral Candida risk factors in humans were documented essentially using data from studies conducted in other areas, hence there is need for comparative and complementary information from the South eastern part of Nigeria. Method: 200 subjects of all age groups of both sexes were randomly examined,by swabbing their palatine mucosa and dorsal tongue with sterile cotton wool,then cultured into Sabouraud dextrose agar plates supplemented with antibiotics and incubated aerobically at 37 degree for 48 hrs. Identification of Candida albicans was done by germ tubes tests, chlamydospores production on cornmeal agar supplemented with 1% Tween 80.Sugar and nitrogen assimilation test using API 20C Auxanogram and potassium nitrate agar. Results: Out of 30 samples that were positive for candida, 15 (50%) were candida albicans. Using the anova test (P < 0.05) this variation is significant (P = 0016). followed by C. dublinensis 3 (13%), C. tropicalis 3 (10%), C. pseudotropicalis 3 (10%), C, glabrata 2 (7%), C. parapsilosis 2 (7%) and lastly C. krusei 1 (3%).However, 53% of the patients were female while 47% were male. Among the HIV positive isolates.67% were HIV isolates not on drugs while 33% positives isolates were on drugs and the percentages of candida species in these patients were as follows C. albicans were 45% followed by C. glabrata and C.tropicalis which were 17% each, C.parapsilosis, C.dubliensis and C.pseudotropicalis were all 8% each. Conclusion: Oral Candidiasis is a marker of systemic diseases and in some cases, it may be the first clinical presentation. There is need for more intensive clinical and laboratory monitoring and possible early intervention to prevent the reoccurrence and resistance to treatment.

Keywords: oral cavity, Candida species, oral Candidiasis, risk factors

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Authors: M. H. Nahaisi, N. M. Almaroum

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Laban is a Libyan traditional fermented milk product. This lactic fermentation has been known in many cities of Libya long time ago as stable, nutritious, refreshing drink especially during the summer. 16 naturally fermented milk samples were collected from different cities located in North West of Libya. The average pH, titratable acidity, fat and total solids were 4.16, 0.73%, 1.54% and 8.12 % respectively. Coliform, yeast and mold counts were 21×10⁴, 39×10⁴ and 41 ×10³ cfu/ ml. respectively. The average Lactococcus, Streptococcus, Mesophilic Lactobacillus / Leuconostoc and Thermophilic Lactobacillus counts were 99 ×10⁷, 96 ×10⁷, 93 ×10⁷ and 15 ×10⁷ cfu / ml. respectively. A total of one hundred forty two lactic acid bacteria (LAB) isolates were identified to the genus level as Lactobacillus (48.59%), Lactococcus (43.66%), Streptococcus (4.93%) and Leuconostoc (2.82%). Sugar fermentation tests have revealed that the most frequently Lactobacillus species was found to be Lactobacillus delbrueckii ssp. lactis (62.32%) followed by Lactobacillus plantarum (31.88%). Furthermore, other selected LAB isolates were identified by API 50 CH test as Lactococcus lactis ssp. lactics, Lactobacillus pentosus, Lactobacillus brevis and Leuconostoc mesenteroides ssp. cremoris.

Keywords: traditional fermented milk, laban, lactococcus, streptococcus, mesophilic lactobacillus, thermophilic lactobacillus counts

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Authors: Harison Masih, Jyotsna Kiran Peter, Sundara Singh, Geetha Singh

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The present investigation deals with diversity of dermatophytes and keratinophilic fungi from different tourist spots such as Agra Fort, Akbar tomb, It-Mat-Ud-Daulah, Mariam tomb, Radha Swami Bagh, and Taj Mahal of Agra City. These fungi are medically important which causes various infections and diseases in humans and animals. The main reservoir of these pathogens are the keratinous substances that increases due to birds and animal activities in the vicinity of monuments, where thousands (5413266) annual visitors from all over the world are visiting. The soil samples were subjected to isolate the pathogenic fungi through bait technique (buffalo skin, chicken feathers, human hair and goat tail hair). Baits were spread over the soil samples and incubated at room temperature for 30-35 days and pure culture isolates were maintained in SDA medium, stored at 4°C. Highest number of visitors were (3906453) from Taj Mahal, minimum 10785 at Mariam tomb annually, the total 271 isolates were encountered from soil samples out of these 18 genera and 38 species were found in different season. Highest incidence was 4.79% frequency shown by Chrysosporium keratinophilum while least 738% frequency occurrence by Trichophyton simii in soil samples. From the present study it was concluded that the incidence of pathogenic fungal isolates were the common in tourists soil that are etiological agents of superficial mycosis. Thus, both human and animal activity seemed to play an important role in occurrence and distribution of keratinophilic and related dermatophytes at various tourist places of Agra city.

Keywords: dermatophytic fungal diversity, bait technique, visitors at tourist spots, human and animal activities, soil samples

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Authors: Essam A. Makky, Chan Cai Wen, Muna Jalal, Mashitah M. Yusoff

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Termites are considered as important pests that could cause severe wood damage and economic losses in urban, agriculture and forest of Malaysia. The ability of termites to degrade cellulose depends on association of gut cellulolytic microflora or better known as mutual symbionts. With the idea of disrupting the mutual symbiotic association, better pest control practices can be attained. This study is aimed to isolate cellulolytic bacteria from the gut of termites and carry out antibacterial studies for the termite. Confirmation of cellulase activity is done by qualitative and quantitative methods. Impacts of antibiotics and their combinations, as well as heavy metals and disinfectants, are conducted by using disc diffusion method. Effective antibacterial agents are then subjected for termite treatment to study the effectiveness of the agents as termiticides. 24 cellulolytic bacteria are isolated, purified and screened from the gut of termites. All isolates were identified as Gram-negative with either rod or cocci in shape. For antibacterial studies result, isolates were found to be 100% sensitive to 4 antibiotics (rifampicin, tetracycline, gentamycin, and neomycin), 2 heavy metals (cadmium and mercury) and 3 disinfectants (lactic acid, formalin, and hydrogen peroxide). 22 out of 36 antibiotic combinations showed synergistic effect while 15 antibiotic combinations showed an antagonistic effect on isolates. The 2 heavy metals and 3 disinfectants that showed 100% effectiveness, as well as 22 antibiotic combinations, that showed synergistic effect were used for termite control. Among the 27 selected antibacterial agents, 12 of them were found to be effective to kill all the termites within 1 to 6 days. Mercury, lactic acid, formalin and hydrogen peroxide were found to be the most effective termiticides in which all termites were killed within 1 day only. These effective antibacterial agents possess a great potential to be a new application to control the termite pest species in the future.

Keywords: antibacterial, cellulase, termicide, termites

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Authors: Wilailak Siripornadulsil, Siriyanapat Tasaku, Jutamas Buahorm, Surasak Siripornadulsil

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The objectives of this study were to isolate LAB from various sources, dietary supplement, Thai traditional fermented food, and freshwater fish and to characterize their potential as probiotic cultures. Out of 1,558 isolates, 730 were identified as LAB based on isolation on MRS agar supplemented with a bromocresol purple indicator and CaCO3 and gram-positive, catalase and oxidase negative characteristics. Eight isolates showed the potential probiotic properties including tolerance to acid, bile salt and heat, proteolytic, amylolytic and lipolytic activities and oxalate-degrading capability. They all showed the antimicrobial activity against some Gram-negative and Gram-positive pathogenic bacteria. Based on 16S rDNA sequence analysis, they were identified as Enterococcus faecalis BT2 and MG30, Leconostoc mesenteroides SW64 and Pediococcus pentosaceous BD33, CF32, NP6, PS34 and SW5. The health beneficial effects and food safety will be further investigated and developed as a probiotic or protective culture used in Nile tilapia belly flap meat fermentation.

Keywords: probiotic, lactic acid bacteria, pathogen, protective culture

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Authors: Geeta Sharma, Suma Chandra

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Sugarcane is one of the major commercial crop playing roles in agriculture and industrial economy of India. Globally sugarcane is affected by approximately 240 diseases caused by various plant pathogenic organisms. Among them, red rot disease caused by the fungus Colletotrichum falcatum, is one of the most important diseases. In the present investigation, one fungal bioagent of Trichoderma harzianum, Pant Bioagent 1 and one bacterial bioagent Pseudomonas fluorescence, Pant Bioagent 2 (PBAT 1 and PBAT 2, respectively) were tested by dual culture method against the pathogen under laboratory conditions. The effectiveness of biocontrol agents was observed against four isolates of C. falcatum. In the case of PBAT1 maximum percent inhibition of pathogen was recorded in isolated Cf 0238 (61.05%), followed by Cf 09 (60.62%) whereas, minimum percent inhibition was recorded in Cf 3220 (48.55%) and in case of PBAT2 maximum mycelial growth inhibition percent was recorded in Cf 767 (50.50%) followed by Cf 088230(48.83%), whereas minimum percent inhibition was recorded in Cf 08 (40.16%) followed by Cf 0238 (41.83%). The present study showed that these biocontrol agents have the potential of controlling the pathogen and can further be used for the management of red rot disease in field.

Keywords: biocontrol agents, Colletotrichum falcatum, isolates, sugarcane

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Authors: Andreev V. A., Kovalenko T. N., Privolnev V. V., Chernavin A. V., Knyazeva E. R.

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Aim: To evaluate the antagonistic activity of Streptococcus salivarius K12 (SsK12) against ENT and oral cavity infection pathogens (S. pneumoniae, S. pyogenes, S. aureus), gram-negative bacteria (E. coli, P. aeruginosa) and C. albicans. Materials and methods: The probiotic strain SsK12 was isolated from the dietary supplement containing at least 1 × 109 CFU per tablet. The tablet was dissolved in the enrichment broth. The resulting suspension was seeded on 5% blood agar and incubated at 35°C in 4-6% CO2 for 48 hours. The raised culture was identified as Streptococcus salivarius with MALDI-TOF mass spectrometry method. The evaluation of SsK12 antagonistic activity was carried out using a perpendicular streak technique. The daily SsK12 culture was inoculated as heavy streaks with a loop at one side of Petri dish with the Muller-Hinton agar (MHA) and incubated for 24 hours at 350 C in anaerobic conditions. It was supposed that bacteriocins would diffuse over the whole area of the agar media. On the next day S. pneumoniae, S. pyogenes, S. aureus, E. coli, P. aeruginosa and C. albicans clinical isolates were streaked at the clear side of MHA Petri dish. MHA Petri dish inoculated with SsK12 (one part) and with the respective clinical isolates (another part) streaked perpendicularly on the same day was used as the control. Results: There was no growth of S. pyogenes on the Petri dish with SsK12 daily culture; the growth of a few colonies of S. pneumonia was noted. The growth of S. aureus, E. coli, P. aeruginosa and C. albicans was noted along the inoculated streak. On the control Petri dish with simultaneous inoculating of the SsK12 strain and the test cultures, the growth of all the testes isolates was noted. Conclusions: (1) SsK12 possesses perfect antagonistic activity against S. pyogenes and good activity against S. pneumoniae. (2) There was no antagonistic activity of SsK12 against S. aureus, E. coli, P. aeruginosa and C. albicans. (3) SsK12 antagonistic properties make it possible to use this probiotic strain for prophylaxis of recurrent ENT infections.

Keywords: probiotics, SsK12, streptococcus salivarius K12, antagonistic activity

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Authors: Enny Suswati, Supangat Supangat

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Background. Ready-to-eat food products are becoming increasingly popular because consumers are increasingly busy, competitive, and changing lifestyles. Examples of ready-to-eat foods include crispy fried chicken. Escherichia coli is one of the most important causes of food-borne diseases and the most frequent antibiotic-resistant pathogen globally. This study assessed the prevalence and antibiotic resistance profile of E. coli from ready-to-eat crispy fried chicken in Jember city, Indonesia. Methodology. This cross-sectional study was conducted from November 2020 to April 2021 by collecting 81crispy fried chicken samples from 27 food stalls in campus area using a simple random sampling method. Isolation and determination of E. coli use were performed by conventional culture method. An antibiotic susceptibility test was conducted using Kirby Bauer disk diffusion method on the Mueller–Hinton agar. Result. Out of 81crispy fried chicken samples, 77 (95.06%) were positive for E. coli. High E. coli drug resistance was observed on ampicillin, amoxicillin (100%) followed by cefixime (98.72%), erythromycin (97.59%), sulfamethoxazole (93.59%), azithromicin (83.33%), cefotaxime (78.28%), choramphenicol (75.64%), and cefixime (74.36%). On the other hand, there was the highest susceptibility for ciprofloxacin (64.10%). The multiple antibiotic resistance indexes of E. coli isolates varied from 0.4 to 1. The predominant antimicrobial resistance profiles of E. coli were CfmCroAmlAmpAzmCtxSxtCE (n=17), CfmCroAmlCipAmpAzmCtxSxtCE (n=16), and CfmAmlAmpAzmCtxSxtCE (n=5), respectively. Multidrug resistance was also found in the isolates' 76/77 (98.70%). Conclusion. The resistance pattern CfmCroAmlAmpAzmCtxSxtCE was the most common among the E. coli isolates, with 17 showing it. The multiple antibiotic index (MAR index) ranged from 0.4 to 1. Hygienic measures should be rigorously implemented and monitoring resistance of E. coli is required to reduce the risks related to the emergence of multi-resistant bacteria

Keywords: antibacterial drug, ready to eat, crispy fried chicken, escherichia coli

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Authors: Çiğdem Sezer, Aksem Aksoy, Leyla Vatansever

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This study has been done for the purpose of evaluation of public health and identifying of enterotoxigenic Staphyloccocus aureus in kitchen of catering. In the kitchen of catering, samples have been taken by swabs from surface of equipments which are in the salad section, meat section and bakery section. Samples have been investigated with classical cultural methods in terms of Staphyloccocus aureus. Therefore, as a 10x10 cm area was identified (salad, cutting and chopping surfaces, knives, meat grinder, meat chopping surface) samples have been taken with sterile swabs with helping FTS from this area. In total, 50 samples were obtained. In aseptic conditions, Baird-Parker agar (with egg yolk tellurite) surface was seeded with swabs. After 24-48 hours of incubation at 37°C, the black colonies with 1-1.5 mm diameter and which are surrounded by a zone indicating lecithinase activity were identified as S. aureus after applying Gram staining, catalase, coagulase, glucose and mannitol fermentation and termonuclease tests. Genotypic characterization (Staphylococcus genus and S.aureus species spesific) of isolates was performed by PCR. The ELISA test was applied to the isolates for the identification of staphylococcal enterotoxins (SET) A, B, C, D, E in bacterial cultures. Measurements were taken at 450 nm in an ELISA reader using an Ridascreen-Total set ELISA test kit (r-biopharm R4105-Enterotoxin A, B, C, D, E). The results were calculated according to the manufacturer’s instructions. A total of 50 samples of 97 S. aureus was isolated. This number has been identified as 60 with PCR analysis. According to ELISA test, only 1 of 60 isolates were found to be enterotoxigenic. Enterotoxigenic strains were identified from the surface of salad chopping and cutting. In the kitchen of catering, S. aureus identification indicates a significant source of contamination. Especially, in raw consumed salad preparation phase of contamination is very important. This food can be a potential source of food-borne poisoning their terms, and they pose a significant risk to consumers have been identified.

Keywords: Staphylococcus aureus, enterotoxin, catering, kitchen, health

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Authors: John Onolame Unuofin, Uchechukuw U. Nwodo, Anthony I. Okoh

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Laccase is constantly gaining status as important biocatalyst in biotechnology. The illimitable potential of its industrial applications and the corresponding aggressive need for phenomenal volumes of extracellularly secreted laccases have called for its interminable production from sources which are able to meet this demand within a relatively short period of time, preferably bacteria. In response to this call, this study was designed to source for laccase-producing bacteria from different environmental matrices. Three sampling environments were chosen such as wastewater treatment plants, University of Fort Hare vicinity and the Hogback woodland, all within the Eastern Cape, South Africa. Samples such as effluents, sediments, leaf litters, degrading wood and rock scrapings were selectively enriched with some model aromatic compounds and were further screened qualitatively and quantitatively on five phenolic substrates ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), Guaiacol, 1-Naphthol, Potassium Ferric Cyanide and Syringaldazine). Basis for selection was their ability to elicit a colour change on at least three of the above mentioned agar based assay substrates. The choice isolates were further identified based on 16S rRNA molecular identification techniques. 33 isolates were screened out of the 40 representative distinct colonies during the qualitative plate screens, while quantitative screens selected out 11 bacterial isolates. They were, based on molecular identification, desginated as members of the genera Pseudomonas, Stenotrophomonas and Citrobacter of the gammaproteobacteria and Bordetalla and Achromobacter of the betaproteobacteria respectively. We therefore conclude based on our outcomes that we may have isolated efficient laccase-producing bacteria, which might be of beneficial significance in catalysis and biotechnology.

Keywords: beta proteobacteria, catalysis, gammaproteobacteria, laccase

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Authors: Naheed Niaz, Nimra Naeem, Bushra Uzair, Riffat Tahira

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Antibacterial activity of different traditional plants essential oils against clinical isolates of Methicillin-resistant Staphylococcus aureus (MRSA) through disk diffusion method was evaluated. All the tested essential oils, in different concentrations, inhibited growth of S. aureus to varying degrees. Cinnamon and Thyme essential oils were observed to be the “best” against test pathogen. Even at lowest concentration of these essential oils i.e. 25 µl/ml, clear zone of inhibition was recorded 9+0.085mm and 8+0.051mm respectively, and at higher concentrations there was a total reduction in growth of MRSA. The study also focused on analyzing the synergistic effects of essential oils in combination with amoxicillin. Results showed that oregano and pennyroyal mint essential oils which were not very effective alone turned out to be strong synergistic enhancers. The activity increased with increase in concentration of the essential oils. It may be concluded from present results that cinnamon and thyme essential oils could be used as potential antimicrobial source for the treatment of infections caused by Methicillin-resistant Staphylococcus aureus (MRSA).

Keywords: Staphylococcus aureus, essential oils, antibiotics, combination therapy, minimum inhibitory concentration

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Authors: Jayson Wau, David Timi, Anthony Harakuwe, Bruce Bowden, Cherie Motti, Harry Sakulas, Rag Gubag-Sipou

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The latex of F. botryocarpa fruit is applied on sores, wounds and other skin infections in Papua New Guinea ethnotherapeutic practices. Systematic bioassay guided separation and isolation of subsequent fractions of latex extracts resulted in three bioactive fractions active against Staphylococcus aureus and Escherichia coli. This study reports structural elucidation of the three isolates. Structures were determined by physical (M.pt and Rf values) and spectroscopic (1D-1H NMR, 2D-HSQC NMR, 2D-HMBC NMR) and MS ESI-POS. The two methylene protons (2H-1) and (2H-3) resonate as triplets at δ 3.59 and δ 4.99 respectively. Electron dense δ 4.99 (2H-3) on (C-3) depicts the strong electron-withdrawing component, quaternary nitrogen (=N= +). Protons resonating at δ 3.88 and 3.89 are singlets depicting two methoxy groups. Both δ 3.88 and δ 3.89 are para-aryls substituents. The methines δ 9.13 and 8.60 are singlets depicting two lone protons on the indolizidinium aryl component. All isolates, (1), (2) and (3) were identified to be ficuseptine by comparing 1D-NMR assignments. 2D-NMR and MS of (2) found it to be ficuseptine chloride '2, 3-dihydro-6, 8-bis (4-methoxyphenyl)-, 1H-indolizinium chloride'. Their counter ions of the ficuseptines were not established and provide promising lead for the further investigation.

Keywords: Ficus botryocarpa, antimicrobial activity, ficuseptine, sores

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Authors: S. Ibrahim, U. B. Abubakar, S. Danbirni, A. Usman, F. M. Ballah, C. A. Kudi, L. Lawson, G. H. Abdulrazak, I. A. Abdulkadir

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Performing culture and characterization of mycobacteria in low resource settings like Nigeria is a very difficult task to undertake because of the very few and limited laboratories carrying out such an experiment; this is a largely due to stringent and laborious nature of the tests. Hence, a rapid, simple and accurate test for characterization is needed. The “SD BIOLINE TB Ag MPT 64 Rapid ®” is a simple and rapid immunochromatographic test used in differentiating Mycobacteria into Mycobacterium tuberculosis (NTM). The 100 sputa were obtained from patients suspected to be infected with tuberculosis and presented themselves to hospitals for check-up and treatment were involved in the study. The samples were cultured in a class III Biosafety cabinet and level III biosafety practices were followed. Forty isolates were obtained from the cultured sputa, and there were identified as Acid-fast bacilli (AFB) using Zeihl-Neelsen acid-fast stain. All the isolates (AFB positive) were then subjected to the SD BIOLINE Analyses. A total of 31 (77.5%) were characterized as MTBC, while nine (22.5%) were NTM. The total turnaround time for the rapid assay was just 30 minutes as compared to a few days of phenotypic and genotypic method. It was simple, rapid and reliable test to differentiate MTBC from NTM.

Keywords: culture, mycobacteria, non tuberculous mycobacterium, SD Bioline

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Authors: Baharuddin Patandjengi, A. Pabborong, T. Kuswinanti

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Bacterial ring rot caused by a gram-positive Coryneform bacterium Corynebacterium michiganensis subsp. sepedonicus is an important disease on potato crops in the world. The disease still belongs to an A1 quarantine pathogen in Indonesia, although it was found in West Java since 2013. The objective of this study was to know the presence of bacterial ring rot in four potato district areas in South Sulawesi. Infected samples were collected from potato fields and storage warehouses in Enrekang, Gowa, Jeneponto and Bantaeng districts. Potato tuber samples were cut and observed their vasiculer vessels and the bacterial ooze was used for isolation on Nutrient Agar and Nutrient Broth–Yeast extract medium. Bacterial isolates were then morphologically and physiologically characterized. A patogenicity test on eggplant and molecular characterization using PCR with specific primer for Cms (50F and Cms 50 R) was revealed for further identification. The results showed that Cms has become widespread in four districts of South Sulawesi. The bacterial ringrot disease incidence in these districts was reached above 30 %. All of 14 bacterial isolates that identified before using standard methods of EPPO, showed DNA band in size of 224 bp in PCR test, which indicated positively belong to C. michiganensis subsp. sepedonicus.

Keywords: bacterial ring rot, clavibacter michiganensis pv. sepedonicus, PCR, potato

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Authors: Abdul Matin, Muazzama Akhtar, Shahid Nisar, Saddaf Mazzar, Umer Rashid

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Antibiotic resistance is an increasing concern worldwide now a day. Neisseria gonorrhea and Staphylococcus aureus are known to cause major human sexually transmitted and respiratory diseases respectively. Nanotechnology is an emerging discipline and its application in various fields especially in medical sciences is gigantic. In the present study, we synthesized multi-walled carbon nanotubes (MWNTs) using acid oxidation method and solubilized MWNTs were with length predominantly >500 nm and diameters ranging from 40 to 50 nm. The locally synthesized MWNTs were used against gram positive and negative bacteria to determine their impact on bacterial growth. Clinical isolates of Neisseria gonorrhea (isolate: 4C-11) and Staphylococcus aureus (isolate: 38541) were obtained from local hospital and normally cultured in LB broth at 37°C. Both clinical strains can be obtained on request from University of Gujarat. Spectophometric assay was performed to determine the impact of MWNTs on bacterial growth in vitro. To determine the effect of MWTNs on test organisms, various concentration of MWNTs were used and recorded observation on various time intervals to understand the growth inhibition pattern. Our results demonstrated that MWNTs exhibited toxic effects to Staphylococcus aureus while showed very limited growth inhibition to Neisseria gonorrhea, which suggests the resistant potential of Neisseria against nanoparticles. Our results clearly demonstrate the gradual decrease in bacterial numbers with passage of time when compared with control. Maximum bacterial inhibition was observed at maximum concentration (50 µg/ml). Our future work will include further characterization and mode of action of our locally synthesized MWNTs. In conclusion, we investigated and reported for the first time the inhibitory potential of locally synthesized MWNTs on local clinical isolates of Staphylococcus aureus and Neisseria gonorrhea.

Keywords: antibacterial activity, multi walled carbon nanotubes, Neisseria gonorrhea, spectrophotometer assay, Staphylococcus aureus

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Authors: Pukhrambam Helena Chanu, Janardan Yadav

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This study investigates the potential of various fungal isolates to solubilize zinc and counteract rice pathogens, with the aim of mitigating zinc deficiency and disease prevalence in rice farming. Soil samples from the rhizosphere were collected, and zinc-solubilizing fungi were isolated and purified. Molecular analysis identified Talaromyces sp, Talaromyces versatilis, Talaromyces pinophilus, and Aspergillus terreus as effective zinc solubilizers. Through qualitative and quantitative assessments, it was observed that solubilization efficiencies varied among the isolates over time, with Talaromyces versatilis displaying the highest capacity for solubilization. This variability in solubilization rates may be attributed to differences in fungal metabolic activity and their ability to produce organic acids that facilitate zinc release from insoluble sources in the soil. In inhibition assays against rice pathogens, the fungal isolates exhibited antagonistic properties, with Talaromyces versatilis demonstrating the most significant inhibition rates. This antagonistic activity may be linked to the production of secondary metabolites, such as antibiotics or lytic enzymes by fungi, which inhibit the growth of rice pathogens. The ability of Talaromyces versatilis to outperform other isolates in both zinc solubilization and pathogen inhibition highlights its potential as a multifunctional biocontrol agent in rice cultivation systems. These findings emphasize the potential of fungi as natural solutions for enhancing zinc uptake and managing diseases in rice cultivation. Utilizing indigenous zinc-solubilizing fungi offers a sustainable and environmentally friendly approach to addressing zinc deficiency in soils, reducing the need for chemical fertilizers. Moreover, harnessing the antagonistic activity of these fungi can contribute to integrated disease management strategies, minimizing reliance on synthetic pesticides and promoting ecological balance in agroecosystems. Additionally, the study included the evaluation of dipping time under different concentrations, viz.,10 ppm, 20 ppm, and 30 ppm of biosynthesized nano ZnO on rice seedlings. This investigation aimed to optimize the application of nano ZnO for efficient zinc uptake by rice plants while minimizing potential risks associated with excessive nanoparticle exposure. Evaluating the effects of varying concentrations and dipping durations provides valuable insights into the safe and effective utilization of nano ZnO as a micronutrient supplement in rice farming practices.

Keywords: biosynthesized nano ZnO, rice, root dipping, zinc solubilizing fungi.

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Authors: Ayman El Behiry, Rasha Nabil Zahran, Reda Tarabees, Eman Marzouk, Musaad Al-Dubaib

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Mastitis is one of the most economic disease affecting dairy cows worldwide. Its classic diagnosis using bacterial culture and biochemical findings is a difficult and prolonged method. In this research, using of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) permitted identification of different microorganisms with high accuracy and rapidity (only 24 hours for microbial growth and analysis). During the application of MALDI-TOF MS, one hundred twenty strains of Staphylococcus and Streptococcus species isolated from milk of cows affected by clinical and subclinical mastitis were identified, and the results were compared with those obtained by traditional methods as API and VITEK 2 Systems. 37 of totality 39 strains (~95%) of Staphylococcus aureus (S. aureus) were exactly detected by MALDI TOF MS and then confirmed by a nuc-based PCR technique, whereas accurate identification was observed in 100% (50 isolates) of the coagulase negative staphylococci (CNS) and Streptococcus agalactiae (31 isolates). In brief, our results demonstrated that MALDI-TOF MS is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology.

Keywords: identification, mastitis pathogens, mass spectral, phenotypical

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Authors: M. Kazou, A. Gavriil, O. Kalagkatsi, T. Paschos, E. Tsakalidou

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Kopanisti cheese is a Greek soft Protected Designation of Origin (PDO) cheese made of raw cow, sheep or goat milk, or mixtures of them, with similar organoleptic characteristics to that of Roquefort cheese. Traditional manufacturing of Kopanisti cheese is limited in small-scale dairies, without the addition of starter cultures. Instead, an amount of over-mature Kopanisti cheese, called Mana Kopanisti, is used to initiate ripening. Therefore, the selection of proper starter cultures and the understanding of the contribution of various microbial groups to its overall quality is crucial for the production of a high-quality final product with standardized organoleptic and physicochemical characteristics. Taking the above into account, the aim of the present study was the investigation of Kopanisti cheese microbiota and its role in cheese quality. For this purpose, four different types of Kopanisti were produced in triplicates, all with pasteurized cow milk, with the addition of (A) the typical mesophilic species Lactococcus lactis and Lactobacillus paracasei used as starters in the production of soft spread cheeses, (B) strains of Lactobacillus acidipiscis and Lactobacillus rennini previously isolated from Kopanisti and Mana Kopanisti, (C) all the species from (A) and (B) as inoculum, and finally (D) the species from (A) and Mana Kopanisti. Physicochemical and microbiological analysis was performed for milk and cheese samples during ripening. Enumeration was performed for major groups of lactic acid bacteria (LAB), total mesophilic bacteria, yeasts as well as hygiene indicator microorganisms. Bacterial isolates from all the different LAB groups, apart from enterococci, alongside yeasts isolates, were initially grouped using repetitive sequence-based polymerase chain reaction (rep-PCR) and then identified at the species level using 16S rRNA gene and internal transcribed spacer (ITS) DNA region sequencing, respectively. Sensory evaluation was also performed for final cheese samples at the end of the ripening period (35 days). Based on the results of the classical microbiological analysis, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, ranged between 7 and 10 log colony forming unit (CFU) g⁻¹, phychrotrophic bacteria, and yeast extract glucose chloramphenicol (YGC) isolates between 4 and 8 log CFU g⁻¹, while coliforms and enterococci up to 2 log CFU g⁻¹ throughout ripening in cheese samples A, C and D. In contrast, in cheese sample B, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, phychrotrophic bacteria, and YGC isolates ranged between 0 and 10 log CFU g⁻¹ and coliforms and enterococci up to 2 log CFU g⁻¹. Although the microbial counts were not that different among samples, identification of the bacterial and yeasts isolates revealed the complex microbial community structure present in each cheese sample. Differences in the physicochemical characteristics among the cheese samples were also observed, with pH ranging from 4.3 to 5.3 and moisture from 49.6 to 58.0 % in the final cheese products. Interestingly, the sensory evaluation also revealed differences among samples, with cheese sample B ranking first based on the total score. Overall, the combination of these analyses highlighted the impact of different starter cultures on the Kopanisti microbiota as well as on the physicochemical and sensory characteristics of the final product.

Keywords: Kopanisti cheese, microbiota, classical microbiological analysis, physicochemical analysis

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Authors: Gachara Grace, Kebira Anthony, Harvey Jagger, Wainaina James

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Aflatoxin contamination of maize in Africa poses a major threat to food security and the health of many African people. In Kenya, aflatoxin contamination of maize is high due to the environmental, agricultural and socio-economic factors. Many studies have been conducted to understand the scope of the problem, especially at pre-harvest level. This research was carried out to gather scientific information on the fungi population, diversity and aflatoxin level during the post-harvest period. The study was conducted in three geographical locations of; Kitui, Kitale and Nakuru. Samples were collected from storage structures of farmers and transported to the Biosciences eastern and central Africa (BecA), International Livestock and Research Institute (ILRI) hub laboratories. Mycoflora was recovered using the direct plating method. A total of five fungal genera (Aspergillus, Penicillium, Fusarium, Rhizopus and Bssyochlamys spp.) were isolated from the stored maize samples. The most common fungal species that were isolated from the three study sites included A. flavus at 82.03% followed by A.niger and F.solani at 49% and 26% respectively. The aflatoxin producing fungi A. flavus was recovered in 82.03% of the samples. Aflatoxin levels were analysed on both the maize samples and in vitro. Most of the A. flavus isolates recorded a high level of aflatoxin when they were analysed for presence of aflatoxin B1 using ELISA. In Kitui, all the samples (100%) had aflatoxin levels above 10ppb with a total aflatoxin mean of 219.2ppb. In Kitale, only 3 samples (n=39) had their aflatoxin levels less than 10ppb while in Nakuru, the total aflatoxin mean level of this region was 239.7ppb. When individual samples were analysed using Vicam fluorometer method, aflatoxin analysis revealed that most of the samples (58.4%) had been contaminated. The means were significantly different (p=0.00<0.05) in all the three locations. Genetic relationships of A. flavus isolates were determined using 13 Simple Sequence Repeats (SSRs) markers. The results were used to generate a phylogenetic tree using DARwin5 software program. A total of 5 distinct clusters were revealed among the genotypes. The isolates appeared to cluster separately according to the geographical locations. Principal Coordinates Analysis (PCoA) of the genetic distances among the 91 A. flavus isolates explained over 50.3% of the total variation when two coordinates were used to cluster the isolates. Analysis of Molecular Variance (AMOVA) showed a high variation of 87% within populations and 13% among populations. This research has shown that A. flavus is the main fungal species infecting maize grains in Kenya. The influence of aflatoxins on human populations in Kenya demonstrates a clear need for tools to manage contamination of locally produced maize. Food basket surveys for aflatoxin contamination should be conducted on a regular basis. This would assist in obtaining reliable data on aflatoxin incidence in different food crops. This would go a long way in defining control strategies for this menace.

Keywords: aflatoxin, Aspergillus flavus, genotyping, Kenya

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Authors: Andi Aliah Hidayani, Asmi Citra Malina A. R. Tassakka, Andi Parenrengi

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Vanname Shrimp is one of the high yielding varieties that are more resistant to virus attacks. However, now this shrimp more death due to virus attack such as white spot disease caused by white spot syndrome virus (WSSV). Various efforts have done to prevent the disease, like immunostimulatory, probiotics, and vaccine. White spot syndrome virus (WSSV) envelope protein VP19 gene is important because of its involvement in the system infection of shrimp. This study aimed to isolate and characterize an envelope protein VP19 – encoding gene of WSSV using WSSV infected Vanname Shrimp sample from some areas in South Sulawesi (Pangkep, Barru and Pinrang). The genomic of DNA were isolated from shrimp muscle using DTAB-CTAB method. Isolation of gene encoding envelope protein VP19 WSSV ws successfully performed with the results of the length of DNA fragment was 387 bp. The results of homology analysis using BLASTn homology suggested that these isolates genes from Barru, Pangkep and Pinrang have closest relationship with isolates from Mexican.

Keywords: vanname, shrimp, WSSV, viral protein 19

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Authors: Rupinder Bakshi, Geeta Walia, Anita Gupta

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Introduction: Urinary tract infections (UTI) are considered to be one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to widespread usage of uncontrolled antibiotics. Materials and Methods: A total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G.M.C Patiala. Urine samples were inoculated on MacConkey’s and blood agar plates by using calibrated loop delivering 0.001 ml of sample and incubated at 37 °C for 24 hrs. The organisms were identified by colony characters, gram’s staining and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby-Bauer disk diffusion method on Muller Hinton agar plates. Results: Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Out of 9149 urine sample, the culture was positive in 25% (2290) samples. Esch. coli was the most common isolate 60.3% (n = 1378) followed by Klebsiella pneumoniae 13.5% (n = 310), Proteus spp. 9% (n = 209), Staphylococcus aureus 7.6 % (n = 173), Pseudomonas aeruginosa 3.7% (n = 84), Citrobacter spp. 3.1 % (70), Staphylococcus saprophyticus 1.8 % (n = 142), Enterococcus faecalis 0.8%(n=19) and Acinetobacter spp. 0.2%(n=5). Gram negative isolates showed higher sensitivity towards, Piperacillin +Tazobactum (67%), Amikacin (80%), Nitrofurantoin (82%), Aztreonam (100%), Imipenem (100%) and Meropenam (100%) while gram positive showed good response towards Netilmicin (69%), Nitrofurantoin (79%), Linezolid (98%), Vancomycin (100%) and Teicoplanin (100%). 465 (23%) isolates were resistant to Penicillins, 1st generation and 2nd generation Cehalosporins which were further tested by double disk approximation test and combined disk method for ESBL production. Out of 465 isolates, 375 were ESBLs consisting of n 264 (70.6%) Esch.coli and 111 (29.4%) Klebsiella pneumoniae. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Conclusion: Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatients UTIs challenging. Ampicillin, Cotrimoxazole, and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help inappropriate and judicious antibiotic usage in a health care setup.

Keywords: Urinary Tract Infection, UTI, antibiotic susceptibility pattern, ESBL

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