Sterile Plastic Containers Research Articles

First report of Botryosphaeria dothidea causing fruit rot on Chinese olive (Canarium album Raeusch.) in Guangdong province of China.

Chinese olive (Canarium album Raeusch.) is a traditional Chinese medicinal plant, mainly cultivated in Guangdong and Fujian provinces in China (Lai et al. 2022). In October 2023, Chinese olive fruit spots were observed in all the Chinese olive orchards surveyed in Chaozhou city (23.75°N, 116.67°E) of Guangdong, with an incidence up to 15%. Early disease symptoms on fruits appeared as circular or irregular, dark brown to black spots with yellowish lesions, and later the spots slowly coalesced to form large necrotic areas, which seriously affected the fruit marketability. To isolate the causal agent, small pieces (~0.3 mm2) of fruit tissue were excised from the lesion margins, and surface-disinfested with 75% (v/v) ethanol for 1 min, followed by 1% NaClO for 3 min, and rinsed three times with sterile water. The pieces were then placed on potato-dextrose-agar (PDA) and incubated at 27°C. Ultimately, four fungal isolates were obtained with similar morphology phenotypes, colonies initially appeared white with irregular margins and after 4-6 days turned dark gray gradually with dense aerial myceliu. Microscopy revealed conidia were single-celled, hyaline, aseptate, fusiform to subclavate, and measured 18.1-22.5 μm × 6.4-9.3 μm (19.8 × 7.4 m on average, n = 30), which were consistent with those descriptions of Botryosphaeria dothidea (Vasić et al. 2013; Zhang et al. 2023). To further identity the isolates, partial sequences of ribosomal transcribed spacer (ITS), translation elongation factor 1-α (TEF1-α), and β-tubulin (TUB2) genes were amplified using primers ITS1/ITS5, TEF-F/R, TUB2-F/R, respectively (Xu et al., 2023; Hong et al. 2006). The sequences of four isolates were identical, and those of representative strain GDCZ-1 were deposited in GenBank (ITS, OR584295; TEF1-α, OR685157; TUB2, OR685158). Using Neighbor-Joining algorithm, phylogenetic tree based on concatenated sequences of ITS, TEF1-α, and TUB2 showed that GDCZ-1 clustered with B. dothidea. To fulfill Koch's postulates, pathogenicity tests were performed on healthy Chinese olive fruits using the needle-prick inoculation method. The fruits were wounded with a sterile needle at the equatorial area (depth of 3-4 mm), and inoculated with 10 µL of spore suspension (106 /mL). The control fruits were inoculated with sterile water. Inoculated fruits were placed in sterile plastic containers to maintain high relative humidity (almost 100%) and incubated at 27°C. After 4 days, the inoculated fruits showed similar symptoms with those observed in the field infected fruits, while the control remained asymptomatic. Pathogen re-isolated from the inoculated fruits showed identical morphological characteristics to the original isolate GDCZ-1. As far as we know, fruit rot caused by Alternaria alternata has been recently reported on C. album in China (Shao et al. 2024). To our knowledge, this is the first report of B. dothidea causing fruit rot disease on C. album in Guangdong. Our report will provide crucial information for studying the epidemiology and management of this disease.

First Report of a New Species, Chrysofolia galloides sp. nov., Causing Black Spots on Gallnuts of Rhus punjabensis var. sinica in China

In June 2021 and 2022, 30-40% ensiform galls of Rhus punjabensis var. sinica (Diels) Rehd. et Wils. showing similar black spots were observed at a same woodland in Zhuyu town (E 107"42', N 32"10', altitude 560 m), Wanyuan, Southwest China. The lesions were usually dotted, rounded to irregular black or blackish brown spots, and scattered to gather into patches on the gall surfaces; the central areas of the lesions were often depressed. The infected galls often grow slowly or even fall off early. Fresh symptomatic galls were randomly collected in June 2021 (9 galls) and 2022 (13 galls). Tissues from the infected galls (ca. 3×5 mm) were surface-disinfected by 75% ethyl alcohol and 10% sodium hypochlorite (NaOCl), then plated on potato dextrose agar (PDA) at 25°C in the dark for six days. Nine isolates from different galls showed similar colony morphology. Similar dominant fungal colonies were isolated from 44.0% and 38.5% of the infected galls in 2021 and 2022, respectively. After sub-culturing of hyphal tips on PDA and oat meal agar (OA)，the colonies showed cottony-felty, annular, whitish to grey-brown after 10 days. Four isolates, IFRDCC1022 and IFRDCC1023 from 2021, and IFRDCC1024 and IFRDCC1025 from 2022 were randomly selected for molecular and morphological evaluations. Selected isolates were cultured on OA and incubated at 21°C at a 12 h/12 h day/night regime to induce sporulation (Jiang et al. 2020). Pycnidia (up to 500 µm diam.) developed within six days or longer. Conidiogenous cells lined the inner cavity of base, which were hyaline, occasionally brownish, ampulliform, 6-12 × 2-4.5 µm. Conidia were solitary, hyaline, guttulate, elliptic, sometimes subcylindric or ovoid, straight to curved, aseptate, (3.5-) 4-6.5 (-8) × 2-3.5 (-4) µm (n=60). Respective DNA fragments were amplified and sequenced with primers ITS1/ITS4 for ITS, LR0R/LR5 for LSU, EF1-728F (Carbone and Kohn 1999) /EF-2 (O’Donnell et al. 1998) for tef1-α, and Bt2a/ Bt2b for tub2 (Glass and Donaldson 1995). The obtained sequences were deposited in GenBank (accession Nos. ITS: OR363211 - OR363214, LSU: OR363215 - OR363218, tef1-α: OR344509 - OR344512, and tub2: OR344513 - OR344516). The morphological characteristics of the new isolates were consistent with the genus Chrysofolia (Crous et al. 2015). The conidiogenous cells (6-12 × 2-4.5 µm) are larger than those of C. colombiana Crous, Rodas & M.J. Wingf. 2015 (5–8 × 2–3 μm) and C. barringtoniae Suwannar., Kumla & Lumyong 2016 (3–7.5 × 2–3 μm), while shorter and wider than those of C. coriariae S.Y. Wang, Yong Wang bis, and Y. Li 2022 (5–20 × 1–3.5 μm). Maximum likelihood (ML), Maximum parsimony (MP) and Bayesian phylogenetic trees generated from the datasets all showed similar topologies, four strains formed a well-supported clade (BS = 99-100%, PP=0.99-1) in Chrysofolia genus, and clearly separated from the other three Chrysofolia spp. and clustered as sister to C. coriariae with high supported values (BS = 97-100%, PP=0.97-1). Based on morphological characteristics and DNA phylogeny, the new isolates were described as a new species, Chrysofolia galloides T. Ma & Z. X. Yang, sp. nov. (Mycobank: MB850312). The two isolates from 2021 (IFRDCC1022 and IFRDCC1023) were randomly selected for pathogenicity tests. Mycelial plugs (ca. 6×6 mm) of a two-week-old colony were placed onto the surface of wounded and unwounded healthy ensiform galls with six or seven replicates. Sterile PDA agar plugs were used as controls. The inoculated galls were incubated at 25°C in sterile plastic containers (21.4×14.0×5.0 cm) lined with wet filter paper for high humidity. The typical symptom appeared within 3 days on 83.3-100% wounded galls, while the unwounded galls and the controls remained asymptomatic. All wounded galls and 20.0-85.7% of unwounded galls showed symptoms within 5 days, while the controls remained asymptomatic. The fungus was reisolated from the lesions and showed identical morphology and 100% sequence similarity of ITS, LSU, tef1-α and tub2 with the initial isolates. No fungus was isolated from the controls. Neofusicoccum parvum is the first fungal pathogen demonstrated to induce brown spots on Chinese gallnuts (Ma and Yang 2022). Therefore, C. galloides is the second fungus confirmed to cause black spots lesions on gallnuts. These findings provide foundations for the comprehensive management of gall production, which have significant economical implication for Chinese gallnut production.

Lowering Cranioplasty Infection Incidence with Novel Bone Flap Storage Protocol: A Retrospective Cohort Study

IntroductionAfter craniectomy, autologous bone flaps may be stored using wet or dry cryopreservation. After brain edema subsides, they are replaced during an operation termed “cranioplasty.” Cranioplasty is associated with 15% infection incidence. MethodsWe conducted a retrospective comparison of infection outcomes between “wet” and “dry” cryopreservation of cranioplasty bone flaps. Historically, bone flaps were stored utilizing “wet” cryopreservation – bone flap storage in 1 L of Lactated Ringer’s solution containing 80mg gentamicin and 2g nafcillin in a sterile plastic container secured in an unsterile plastic bag. Our newer dry cryopreservation protocol involved storage in gauze soaked in 80mg gentamicin and 2g nafcillin within a 3-layer sterile bag system. Results119 autologous bone flaps were included with median follow-up of 3.9 months from cranioplasty. Overall, 10.9% became infected requiring subsequent surgery. 18.4% of 49 bone flaps stored using wet cryopreservation became infected compared to only 5.7% of 70 dry cryopreservation bone flaps (p 0.038; Relative risk 0.311; Absolute risk reduction 12.7%). Tobacco use (p 0.076; RR 3.17) was not associated with increased infection risk. Infection incidence was similar for traumatic craniectomy indications compared to the other indications (12.0% trauma versus 10.1% other; p 0.750). On average, infected cranioplasty patients spent 8.5 more days hospitalized and faced increased risk of additional complications. ConclusionDry cryopreservation significantly decreases infection after cranioplasty when compared with wet cryopreservation, which mitigates additional morbidity, mortality, and costs attributable to cranioplasty infection. Other nonmodifiable risk factors for cranioplasty infection were identified.

EVALUATION OF ANTIBACTERIAL POTENTIAL OF LICORICE AND XYLITOL CONTAINING SUCROSE FREE HARD CANDIES IN HIGH CARIES RISK PATIENTS

Objectives: To evaluate the effi cacy of sucrose-free hard candies containing xylitol and licorice for preventing dental caries in high-risk individuals. Materials and Methods: Total 80 participants were enrolled based on inclusion criteria. Participants were divided at random into four groups of twenty. Group A consumed stevia-sweetened hard candies including licorice extract, Group B consumed stevia-sweetened hard candies containing xylitol extract, Group C consumed stevia-sweetened hard candies, and Group D consumed sucrose-sweetened hard candies. For the fi rst ten days, participants were asked to eat 2 candies each day; saliva samples were collected at Day 0, after fourteen days following the baseline saliva collection, and after six months after the intervention. Each participant was asked to spit into a sterile plastic container until a suffi cient quantity (5ml) of un-stimulated saliva was collected. It took place between 9 and 10 a.m. For at least an hour before the saliva sample was taken, the individual was requested to abstain from all forms of oral care. The statistical analysis was done by SPSS version 20. Results: Eighty participants finish the experiment and follow the instructions. CFU and pH data for streptococcus mutans in saliva were done as experimental analysis. The mean percentage change in CFU of Streptococcus mutans (%) varied signifi cantly across hard candies over the course of each time period (baseline to fourteen days, P<0.001; fourteen days to six months, P<0.001; and baseline to six months, P<0.001). There was a statistically signifi cant difference in mean salivary pH across various assessment periods within Base line to fourteen days (P<0.001) and Fourteen days to six months (P=0.019) and Base line to six months (P<0.001). The connection between bacterial count and salivary pH was found to be -0.142 (P=0.212) using Pearson's correlation coefficient. Conclusion: Licorice or xylitol extract are used to make sugar-free hard candies that are antibacterial, although the effect wears off with time. Hard candies without sugar have been proven to decrease cariogenic bacteria, while having an essentially neutral effect on pH.

Influence of Pepper Additive and Packaging Styles on Nutrient, Fungi and Aflatoxin Compositions of Stored ‘Robo’ (Deffatted Melon Snack)

Robo, an indigenous defatted melon snack commonly consumed in Nigeria generates a percentage of rural cash earnings for its producers. However, studies on the possible aflatoxin contamination of this product due to associated fungi and their control using cost effective measures are scarce. This study demonstrates the production of Robo, reports its nutrient composition, effects of pepper additive, packaging styles, and its possible fungi/aflatoxin contamination. Powdered chilli pepper was supplemented to paste of melon seed at different concentrations; 50g (T1), 100g (T2), 150g (T3) and 200g (T4) during Robo production while those with no pepper additive were treated as control. Thereafter, they were packaged in sterile polyethylene bag, plastic container and white paper and stored at 28±2°C for 12 weeks. The nutrient, fungi and aflatoxin compositions of fresh and stored Robo samples were determined. The result showed that paper-stored Robo samples had the highest carbohydrate (20.53±4.91), crude protein (40.96±3.79), ash (7.47±0.89), fibre (8.90±1.47) and pH (6.81±0.55) and least moisture (8.14±0.58) contents. Most dominantly associated fungi with this food are Aspergillus flavus, A. parasiticus, Fusarium oxysporium, Penicillium sp, Candida albicans, A. ustus, A. niger, Trichoderma sp, Rhizopus stolonifers and A. oryzae. The polyethylene bag-stored samples record the highest fungal incidence (88) followed by plastic-stored samples (81) while the paper stored samples had the least (76). Plastic-stored samples had the highest aflatoxin content AFB1 of 1.02±0.19 while the polythene stored samples had the highest AFB2 (0.45±0.31), AFG1 (0.02±0.41) and AFG2 (0.02±0.02) respectively. However, paper-stored samples had the least AFB1 (0.52±0.29), AFB2 (0.34±0.09), AFG1 (0.00±0.10) and AFG2 (0.01±0.04). Interestingly, it was observed that increased concentrations of chilli pepper from T1-T4 resulted in the corresponding decrease in their aflatoxin compositions. In addition pepper addition improved the organoleptic properties of the Robo samples as T2 was generally accepted after twelve weeks of storage. The implications of these findings are discussed.

Microbial and sensory quality of powdered beverage produced from blends of baobab pulp and moringa leave during storage

The study investigated the microbial and sensory quality of powdered beverages produced from blends of baobab pulp and moringa leave flour. Samples were prepared and stored in sterile plastic containers and bottles, were stored at room temperature 27±2 0c and in the refrigerator for 21days. Sensory evaluation was done using the following parameters, appearance, mouth fill, taste, flavor and general acceptability. The samples most preferred in relation to all the parameters tested is samples A and B. Total aerobic plate count of the bacterial cells for control samples A-E ranged from 5.2×106_ 8.2×106. cfu/ml Samples A-E ranged from 8.5 x 105 - 3.04×107 for plastic container and 1.15×105-1.6×107 for samples stored in bottles at room temperature ,while samples stored in the refrigerator ranged from 5.6 x 105 – 2.3x107 for plastic containers and bottles 4.6 x 106 – 9.5x106 for week one. For week two samples A-E for plastic and bottles ranged from 3.7 x 105 - 8.5×105 and 3.2 ×105 - 9.6 x 106 for samples stored at room temperature and refrigerator respectively. For week three samples A-E stored in plastics and bottles at room temperature and refrigeration ranged from 3.5x105-1.7x106 and 1.0x105 – 3.0 x 105. The bacterial isolates were characterized using morphological, biochemical tests and the identity of the isolates were confirmed with a microgen test kit .The species of bacteria identified includes: Bacillus Spp, Escherica coli, Klebsiella pneumonia, Staphylococcus aureus, Salmonelia typhi and shiegella. From this study it shows that the baobab pulp and moringa leave flour blend beverage should be stored in the refrigerator or consumed immediately after processing in order to maintain its organoleptic properties and avoid microbial contamination.

Source of household water as main risk factor of soil-transmitted helminth infections among elementary school pupils in Wamena District, Jayawijaya Regency, Papua

BackgroundSoil-transmitted helminth (STH) infections are caused by three types of worms: Ascaris lumbricoides, Trichuris trichiura, and hookworms. Soil-transmitted helminth infections have significant health and socioeconomic implications for communities in developing countries, particularly in Indonesia. This study aims to determine the prevalence and risk factors of STH infections among elementary school pupils in Wamena District. MethodsThe study design used was a cross-sectional design involving 317 elementary school pupils. The children were interviewed using a structured questionnaire that included demographic information, hygiene, and environmental sanitation data of the respondents. Stool samples were collected from all consenting participants in sterile plastic containers and were analyzed within 24 hours of collection, using the Kato-Katz method. The data were analyzed using bivariate statistical analysis (Chi-square) and multivariate analysis (logistic regression). ResultsThe prevalence of helminthiasis among elementary school pupils was 19.9% (63/317). STH prevalence of mild category was 17.4% (55/317). The risk factors for STH infections among school children were the habit of not washing hands with soap after defecation, with an odds ratio of 5.04 [95% CI (2.22-11.48)], and the source of water for household use, with an odds ratio of 7.22 [95% CI (3.66-14.22)]. ConclusionThe prevalence of helminthiasis was found to be 19.9% (63/317), with an STH prevalence of mild category. Risk factors for STH infections included the habit of not washing hands with soap after defecation and the source of water for household use.

Isolation and identification of non-saccharomyces ethanol and thermo-tolerant yeasts strains from fermented carbohydrate wastes

Ethanol is an important chemical with its potential as a biofuel to displace the environmentally unfriendly fossil fuels. Isolation and identification of ethanol and thermo-tolerant yeast capable of producing ethanol were carried out in this study. Seventy (70) samples from different fermented foods and agricultural wastes from millets, sorghum, maize, rice and guinea corn were obtained from local sellers in Panteka, Farin-Gida, Barkallahu and Barnawa in Kaduna State (North West), Nigeria were used in this research. The samples were collected using a sterile plastic container and were transported immediately to the Biotechnology laboratory of the Nigerian Defence Academy in iced chamber. They were serially diluted in distilled water and inoculated on yeast extract peptone dextrose agar (YEPD) media plates supplemented with 0.2% chloramphenicol. The plates were incubated at 30°C for 72 hours and developed colonies were sub-cultured to obtain pure culture which was observed for their morphological and microscopic characteristics. After examination, ten (10) yeast isolates were obtained from the seventy (70) samples collected. Flocculation test, Hydrogen sulphide test, preliminary alcohol production and sugar fermentation test were carried out on the isolates and their tolerance to ethanol at varying concentration 5%-20% (v/v) and temperature at 37oC-45oC were studied. Using the internal transcribed spacer (ITS) 1 and 2 regions primer, the DNA were sequenced and the isolates identified. Four isolates FG 17, FG 27, P1 and BN50 showed ethanol tolerance potentials at 20% ethanol and were identified as strains of Diutina rugosa, Candida sp, Candida ethanolica and Candida tropicalis while one isolate FG 35 showed good thermo-tolerance ability at 45oC and was identified as Trichosporon sp.

PHYSICOCHEMICAL AND MICROBIAL ANALYSIS OF BOREHOLE WATER IN HOSPITALS AROUND BENIN CITY, NIGERIA

Ground water is known to be the major source of water in many hospitals, extracted through domestic boreholes for public and private usage. This study was carried out to access the physicochemical and bacteriological quality of borehole water in some hospitals in Benin City. Borehole water samples were collected from three different hospitals (Hospital A, Hospital B and Hospital C) around Benin City, Edo State into sterile plastic containers to avoid additional contamination before being taken to the laboratory for physicochemical and bacteriological assessment. The results showed the presence of microorganisms in the water samples. The highest bacteria heterotrophic counts were seen in hospital C (81.0±0.5), while the lowest was observed in hospital B (7.0±0.5). The probable identified bacterial isolates were Salmonella sp., Staphylococcus sp., Pseudomonas sp., Bacillus sp., Shigella sp., Klebsiella sp. and Escherichia coli. The Physicochemical analysis revealed that the highest pH value (6.06±0.33) was observed in water from hospital A, total suspended solids (1.30±0.08 mg/ml), turbidity (2.73±0.11 NTU), nitrate (1.05±0.05 mg/ml), phosphate (0.23±0.00 mg/ml), sulfate (1.84 mg/ml), biological oxygen demand (0.03±0.01 mg/ml) and chemical oxygen demand (0.81±0.09 mg/ml) were also recorded. No heavy metals were found from the water samples within the three hospitals. This study revealed the occurrence of microorganisms, hence the need for water treatment in hospitals. Environmental agencies should also ensure compliance with relevant standards to avoid risks to human health.

Autologous versus synthetic cranioplasty

Background. Cranioplasty has been described in history as far back as the 16th century. The use of autologous cranioplasty has been published since 1821 and is still under practice today worldwide. Recent evidence however has suggested increased complication and revision rates with the use of autologous bone. We compared our results of autologous cranioplasty versus synthetic material. Methods. A retrospective study was carried out of cranioplasty procedures at our unit between August 2009 and March 2018. Bone flaps were placed in a sterile sealed plastic container and stored at -81 degrees. Swabs and bone chips were used for cultures and bone flap disposed if positive. On re-implantation, the bone was thawed at room temperature and soaked in gentamicin. Synthetic cranioplasties were constructed using thin-slice CT to design a custom flap for each patient. Results. 144 cranioplasties were studied. 51 own bone and 93 synthetic. The average delay in cranioplasty was 286 days (Range 16 – 1264 days). The overall complication rate for all 144 cranioplasties was 20.8%; Autologous 31.4% and synthetic 15.1%; p 0.031. Bone flap infection rate overall for all 144 cases was 9.7% - Autologous 11.8% and Synthetic 8.6%; p 0.565. The revision rate was found to be 13.2% overall; 23.5% for autologous and 7.5% for synthetic. The difference in revision rate was found to be statistically significant (p 0.01). Conclusion. Revision rate and overall complication rate were higher in the own bone group with P<0.05. There was no difference in infection. Our results mirror recent publications and should be considered when undertaking a cranioplasty.

Diversity and Distribution of Fungi Associated with “Ikwe Pond” Water Source: Their Impact on Water Quality and Human Health

Purpose: Fungal infections are becoming more and more important because of increasing numbers of immunosuppressed patients. Nonetheless, waterborne fungi are associated with variety of health related conditions. Fungi in the aquatic ecosystem perform essential functions in biogeochemical cycles. However, knowledge of fungal communities associated to a particular aquatic ecosystem is important. In the present study, we seek to investigate the fungal distribution and diversity in Ikwe Pond, located in Eket.
 Methodology: A total of thirty different water points were sampled using 30 thoroughly washed 0.50L sterile plastic containers and cultured on SDA plates with 1ml of dilution factors 10-3 and 10-6 used as inoculums. Six fungal species namely Candida tropicalis, Aspergillus niger. Cryptococcus sp, Absidia sp, Geotricium sp, were isolated from the 30 sampling points.
 Findings: Upstream recorded a total of 27 spores, midstream had 14 spores while downstream recorded 8 spores. Also the colony count revealed that upstream had the highest number of fungal spores count. Clearly, the upstream (59.3%) and the midstream (78.6%) sampling points were dominated by Aspergillus niger while the downstream water column was dominated by Candida tropicalis (50%).
 Unique Contribution to Theory, Practice and Policy: In many countries, there has been little focus on the occurrence of fungi in water which can lead to severe health related issues but with monitoring on the frequency and distribution of fungi in our waters, this perceived health related complications can be checked.

Milk microflora for mastitis in cows

Mastitis in cows is currently a pathology that causes great economic damage to livestock farms around the world. The quantitative and qualitative indicators of milk are decreasing. Prevention and treatment of breast inflammation in cows is one of the main tasks of veterinarians. For successful treatment, it is necessary to understand the etiology, especially in infectious mastitis. In order to determine the pathogenic microflora that causes mastitis in cattle, we conducted a bacteriological study of 10 milk samples from cows with breast inflammation in one of the livestock farms of the North-Western region. For the bacteriological study of milk, each sample was taken into a sterile disposable plastic container, the first trickles of milk were pre-deposited. As a result of the conducted studies, mastitis in cows caused by different microorganisms is observed, it follows that not only the farm has its own microbial landscape, but in each case different microorganisms cause mastitis. Cultures of conditionally pathogenic microflora - enterobacteria, coccoid microflora – were isolated from the material. Cultures of Salmonella, Pseudomonas aeruginosa, Staphylococcus aureus and other pathogenic microflora were not isolated in any sample. High resistance of staphylococci to antibacterial drugs of different groups was revealed. Based on the data obtained, it is worth noting the need for a bacteriological study before each appointment of mastitis treatment in cattle, especially during antibacterial therapy. It is necessary to take into account the developing resistance in pathogenic and conditionally pathogenic microorganisms. It is also important to note the relevance of creating a drug for the specific prevention of mastitis of cows of bacterial etiology.

Prevalence and Associated Risk Factors of Soil-Transmitted Helminth Infections among Schoolchildren around Lake Tana, Northwest Ethiopia.

Soil-transmitted helminths (STHs) are among the most common neglected tropical diseases widely distributed in tropical countries with poor socioeconomic development. Ascaris lumbricoides, Trichuris trichiura, and hookworm are the three major STHs. This study aimed to determine the prevalence of STHs and associated risk factors among schoolchildren in selected primary schools around Lake Tana, Northwestern Ethiopia. A school-based cross-sectional study was conducted from February to May 2021 involving 337 study participants. A systematic random sampling method was utilized to select the study participants from the selected schools. Data related to sociodemographic characteristics of the study participants and risk factors for STH infections were collected using a pretested questionnaire. Stool samples were collected in sterile plastic containers from each participant and processed using Kato-Katz thick fecal smear and Ritchie's concentration techniques. The data were analyzed using the Statistical Package for Social Sciences software tool version 23, and factors with a p < 0.05 were considered as statistically significant. The overall prevalence of STH infection was 38.3% (95% CI: 33.1-43.7). Hookworm infection was the predominant STH infection, which was detected in 26.1% schoolchildren followed by A. lumbricoides (14.8%) and T. trichiura (1.5%). Most of the study subjects (34.1%) had single infections, whereas only 4.2% study subjects had multiple helminthic infections. Among the risk factors considered in the study, lack of shoe wearing habit (adjusted odds ratio [AOR]=29.5; 95% confidence interval [CI]=6.59-132.55; p < 0.001), lack of knowledge on the prevention and control methods (AOR = 5.41; 95% CI = 2.44-11.98; p < 0.001), engagement in irrigation activities (AOR = 2.14; 95% CI = 1.02-4.57, p = 0.049), lack of toilet (AOR = 3.06; 95% CI = 1.31-7.16; p = 0.01), children grades of 5-8 (AOR = 2.62; 95% CI = 1.26-5.43; p = 0.01), playing on soils (AOR = 5.90; 95% CI = 2.79-12.49; p < 0.001), lack of fingernail trimming habit (AOR = 3.21; 95% CI = 1.57-6.55; p = 0.001), and male gender (AOR = 2.28; 95% CI: 1.19-4.39; p = 0.013) were significant explanatory factors for STH infection among schoolchildren in the study area. The present study showed that STHs were common among schoolchildren around Lake Tana. Therefore, education on personal and environmental hygiene should be taken into account to reduce the prevalence of STH infection in the study area.

The Prevalence of Extended Spectrum Beta-Lactamase Producing Uropathogenic Escherichia coli from Mouau Female Hostel Students

Aim: The study is aimed at determining the antimicrobial susceptibility pattern and production of Extended-spectrum Βeta-Lactamase (ESBLs) among Escherichia coli isolates from urine samples.&#x0D; Methods: A total of 35 samples of early morning mid-stream urine samples of about 10-15 ml were collected from Michael Okpara University of Agriculture Umudike (MOUAU) female hostel students; using sterile plastic containers. Each urine sample was inoculated on MacConkey Agar (MCA). Isolates were identified based on morphological features, Gram staining, and biochemical characterization. The antibiotic susceptibility test was done by the Kirby Bauer Disc diffusion method on Muller Hinton agar. ESBL detection was done as per Clinical and Laboratory Standard Institute (CLSI) guidelines, using the double disc synergy test method.&#x0D; Results: The study reviewed a total of 12 isolates of Escherichia coli of which is (34.29%) of which were obtained from urine samples. The susceptibility test results showed that most isolates were susceptible to Ofloxacin with 83.3% susceptibility, followed by Levofloxacin (66.7%), whereas gentamicin, nalidixic acid, cefuroxime, and cefotaxime exhibited a susceptibility rate of 33.3%. On the other hand, the highest resistance rate was to imipenem (96.7%), followed by Amoxicillin and ampiclox with 83.3%, while the least percentage resistance was observed in Ofloxacin with (8.3%) and Levofloxacin with (16.7%). Extended-spectrum Βeta-Lactamase (ESBLs) production among E. coli isolates was detected in 25.0% of the potential ESBL producers. The result of the multiple antibiotic resistance index showed that the isolate (E20) was significantly higher than other isolates with p&lt;0.05, whereas E3, E7, E10 and E12 had same resistance pattern.&#x0D; Conclusion: So drug resistance due to ESBL production is a severe threat in UTIs, narrowing down the choice of antibiotics for treatment. So, there is a need to introduce routine screening for ESBL production for all uropathogenic Escherichia coli, causing urinary tract infections.

First Record of Alternaria alternata causing necrosis of Thuja (Thuja occidentalis) in Kazakhstan.

Thuja is one of the ornamental plants used for landscaping parks and health resorts. The plant is distinguished by a pyramidal and conical crown shape and the presence of many thin branches with scale-shaped needles, green all year round. In addition, this plant has a number of antimicrobial properties, which affects the popularity of the plant in landscaping the health resort territory (Bakht et al. 2020, Chindyaeva et al. 2020). In January 2020, symptomatic Thuja plants were observed in Southern Kazakhstan. Symptoms included distortion of the crown. External examination of the trees revelaed the presence of white fluffy mycelium on Thuja branches. The branches acquired a yellow color with a necrotic lesion developing below the affected area. Samples of infected branches from different Thuja trees (n = 13) were collected. The infected branches were cut into small pieces (5 × 5 mm), washed in 70% ethanol for 30 min, and then rinsed three times with sterile distilled water. Later, these pieces were placed on Sabouraud's medium (Laboratorios Conda S.A., Spain) and incubated at 28°C for 7 days. Yellow-green colonies grew from the pieces of wood. The colonies had a light gray-whitish aerial mycelium. Conidia (n = 35) were pale to dark brown in color, irregular and ellipsoid to ovoid conical in shape. The size of the conidia varied from 5 to 25 µm × 6 to 12 µm (n = 40) with longitudinal and transverse septations. These morphological characters were previously described and corresponded to the Alternaria alternata (Simmons et al. 2007). Genomic DNA was extracted from mycelium using the liquid nitrogen and phenol-chloroform extraction method (Butler 2012). A 568 bp product of the Alt a1 gene and 472 bp product of the calmodulin protein-coding gene was amplified using following primer pairs Alt-for/Alt-rev (Hong et al. 2005) and CALDF1/CALDR1, respectively (Lawrence et al. 2013) (Integrated DNA Technologies, Inc., Coralville, IA, USA). The PCR reaction was done in a SimpliAmp thermal cycler (Applied biosystems, Waltham, MA, USA) under the following conditions: initial denaturation at 94 °C for 1 min, 35 cycles at 94°C for 30 s to denature, 57°C for 1 min for annealing, and 72°C for 1 min for extension. A final extension step at 72°C for 10 min was also included. The sequencing was done using BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems) and the sequence was deposited in GenBank with accession no. OL542696 calmodulin). These sequences were compared with other sequences in the GenBank by using the BLAST analysis (MZ222274.1 and MN473132.1). The phylogenetic analysis was carried out with MEGA 6 software (The Pennsylvania State University, University Park, PA, USA). To confirm the pathogenicity, 10 thuja branches from healthy trees from another area without visible pathologies were inoculated with a suspension of conidia (100 conidia/ml; obtained from 2-week-old cultures). Control samples were inoculated with sterile distilled water. The inoculated branches were placed in sterile plastic containers to maintain high humidity and incubated for 10 days at 28°C. After 7 days, irregular shaped lesions and fungal growth was observed at the site of inoculation. The affected area gradually increased in size with simioar symptomatology to that described above. Re-isolation of the pathogen and identification based on morphological features and sequencing confirmed the presence of the A. alternata pathogen. To our knowledge, this is the first report of A. alternata causing branches of thuja in Kazakhstan. Thuja is a rare plant species for this region; the cost and care are expensive. This case will allow timely diagnosis of the disease caused by Alternaria spp. in the future. It is necessary to develop preventive measures and a protocol for the treatment of thuja from a fungal infection. Bakht, J., 2020. Antibacterial activity of the crude extracts from medicinally important Thuja occidentalis. Pak J Pharm Sci. 33(2): 627-630. PMID: 32276908. Butler, J.M., 2012. Chapter 2 - DNA extraction methods. pp. 29-47 in Butler JM (Ed) Advanced topics in forensic DNA typing: Methodology. San Diego, Academic Press. doi: 10.1016/C2011-0-04189-3. Chindyaeva., L.N., et al. 2020. Comparative assessment of the phytoncidity of woody plants in the selection of species for landscaping: the possibility of use in sanatorium-and-spa practice. Vopr Kurortol Fizioter Lech Fiz Kult. 97(4): 44-51. Russian. doi: 10.17116/kurort20209704144. Hong, S.G. et al. 2005. Alt a1 allergen homologs from Alternaria and related taxa: analysis of phylogenetic content and secondary structure. Fungal Genet Biol 42:119-129. doi:10.1016/j.fgb.2004.10.009 Lawrence, D.P. et al. 2013. The sections of Alternaria: formalizing species-group concepts. Mycologia 105: 530-546. DOI: 10.3852/12-249. Simmons, E. G., 2007. Alternaria: An Identification Manual. CBS, Fungal Biodiversity Center, Utrecht, Netherlands.

S5.4c Isolation of keratinophilic fungi of the genus Microsporum from the soils of Moscow, Russia

5.4 Free oral paper session, September 22, 2022, 3:00 PM - 4:30 PMIt is reported about the discovery of zoophilic and geophilic dermatophytes in the soils of city parks and adjacent territories of Moscow, Russia. Microscopic fungi of the Microsporum genus were found in 27/50 samples (54%): Microsporum gypseum (42%) and Microsporum canis (12%), their number was higher in the soils taken from the adjacent areas where domestic animals are walked. This increases the risk of infection of animals upon contact with soil.ObjectivesThe research was conducted to determine the frequency of occurrence of potential pathogens of animal dermatophytosis in those areas of the soil cover where pets are walking. Soil samples were taken from several areas: the adjacent territories along the streets of Vishnevsky and Kostyakov, designated as areas 1 and 2, the park ‘Dubki’, Lianozovsky, Timiryazevsky, designated as 3, 4, and 5, respectively.Areas 1 and 2 are narrow patches of soil adjacent to residential buildings and sidewalks where pets are most often walked. Areas 3 and 4 are areas of soil adjacent to park paths, where animal walking is less common. Area 5 is a relatively little-visited place in the forest park, where animals are rarely walked.MethodsBefore sampling, vegetation cover and rubbish were removed from the soil area, then ∼ 100 g of soil was taken from the surface using aseptic rules in sterile plastic containers. A total of 10 soil samples were taken from each plot. The soil was dried at room temperature, sieved to remove rubbish, and plant parts, and placed in sterile Petri dishes.Isolation of fungi was carried out by the ‘bait’ method: samples of sterile pet hair about 1 cm long were placed on the surface of the soil, the soil was moistened with sterile distilled water with the addition of a selective additive for the isolation of dermatophytes (Dermasel, Oxoid). Petri dishes were periodically inspected for fungal growth on the surface of the wool and moistened if necessary.When the growth of microscopic fungi was detected on the surface of the wool, they were sterilely transferred to Petri dishes with wort agar medium (M129, HiMedia) to obtain a pure culture and identification. Only microscopic fungi of the genus Microsporum were taken into account.ResultsAs a result of the research, a total of 27 isolates of microscopic fungi, 21 of Microsporum gypseum and 6 of Microsporum canis were isolated from the soils of Moscow: 7 isolates at the area 1 (6 of M. gypseum, 1 of M. canis), 8 isolates at the area 2 (5 of M. gypseum, 3 of M. canis), 5 isolates at areas 3, and 5 (4 of M. gypseum, 1 of M. canis), 2 isolates of M. gypseum at the area 5. The results are illustrated in Figure 1.ConclusionSoils located in adjacent areas are more polluted by the hair of pets that walk there. Therefore, these soils also contain keratinophilic microscopic fungi, which includes the pathogens of animal dermatophytosis. In such places, animals are susceptible to infection with fungal infections directly from the soil.

EFFECTIVENESS OF BROMELAIN MOUTHRINSE IN GINGIVAL INFLAMMATION AND BACTERIAL PLAQUE AMONG ADOLESCENTS: A RANDOMIZED CLINICAL TRIAL

Background: Mouthwashes are adjunct to regular brushing and flossing. Chlorhexidine is considered as a gold standard among mouth rinses. However, the most commonly known side effects include discolouration of teeth and the tongue particularly on the dorsum part. Hence the study was conducted to evaluate the effectiveness of Bromelain mouth rinse in gingival inflammation and bacterial plaque among adolescents.&#x0D; Methods and methodology: This double-blinded, randomized controlled trial was conducted in accordance with Consolidated Standards of reporting Trials (CONSORT) in Public Health Dentistry Department at Saveetha University. The protocol was approved by the Institutional Review Board (IRB). 30 patients with moderate gingivitis were included in the study. All subjects signed an IRB-approved consent form. Participants from 15 to 19 years are recruited from the Urban health programmes through dental camps.&#x0D; Results: No attempt was made for oral prophylaxis before the commencement of the study. The severity of gingivitis and plaque accumulation were assessed by gingival and plaque indices. Gingival index (Löe and Silness, 1963) and plaque index (Löe, 1967) were measured initially at baseline The trial was followed up for 30 days. The participants were instructed to rinse with plain water thoroughly. They were then directed to spit into a sterile plastic container after 5 minutes.At baseline, there was no significant difference in GI and PI scores among the groups (P=0.78 and 0.62) respectively. At one month follow up, there was significant difference in the mean GI and PI scores (P=0.02 and P=0.01) respectively.&#x0D; Conclusion: Bromelain mouthwash is beneficial in improving gingival status due to its profound styptic action, with sufficient reduction in plaque scores without any adverse effects.&#x0D;

Parasitas gastrintestinais em galinhas de raças ornamentais de um criatório particular na cidade Juiz de Fora, Brasil

To evaluate the occurrence of gastrointestinal parasites in chickens of ornamental breeds from a private farm located in Juiz de Fora, Minas Gerais. Fecal samples were collected from the animals individually, collecting the feces right after defecation. The collections were carried out in September 2018. The samples were packed in sterile plastic containers, identified and sent to the Laboratory of Parasitology and Parasitic Diseases of the Centro Universitário de Valença (UNIFAA), for analysis by the simple centrifuge-float technique. Two genera of helminths, Capillaria spp. and Ascaridia spp., and the protozoan Eimeria spp. It can be concluded that the gastrointestinal parasites found in the ornamental chickens studied did not differ from the endoparasites found in conventional free-range chickens. Coproparasitological exams are an excellent, fast and cheap way to verify/identify parasitic forms, which, once diagnosed, must be fought so that they do not compromise the health of the animals, especially in cases of ornamental birds that, because they are less rustic, may be more sensitive to parasites.

Investigating the Presence of Salmonella spp. in the Feces of Red Deer (Cervus elaphus) in Istanbul

Wildlife is an important source of many infectious agents for domestic animals and humans. Increasing interactions between humans, domestic animals, and wildlife may cause the infection of contagious agents between the species. Domestic animals can be easily infected with Salmonella serovars as a result of its asymptomatic presence in the intestinal flora of wild animals and its spreading around. The presence of Salmonella species in the feces of red deer, which is an important member of wildlife in Turkey, was investigated in the present study. A total of 240 fecal samples were collected in sterile dry plastic containers from free-ranging deer at the Deer Breeding Farm in Istanbul Belgrad Forest, at least once a month for a year. The samples collected were examined through conventional culture and molecular methods (PCR). Salmonella spp. was not isolated from the samples examined.

Effect of Boiling, Oven Drying and Roasting On the Proximate Composition and Microbial Loads of Cane Rat

The study examined the effect of proximate composition, mineral and microbial loads of cane rat (Thryonomys swindarianus) meat of different processing methods. Three (3) live adult cane rats were purchased from reputable cane rat domestication farm in Abeokuta and transported to the wildlife laboratory of the Federal University of Agriculture where the samples were slaughtered and subjected roasting, boiling and oven drying. Small piece of each processed cane rat meat was extracted and placed in sterile plastic containers for proximate, mineral and microbial loads for three (3) consecutive days. The standard procedure described by Holt et al., (1994) method was used for the analysis of moisture content, crude protein, total available carbohydrate and ash. Data obtained were subjected to analysis of variance using least significant difference to separate the means and line graph was used for the microbial count. The results showed that from day 1 to day 3 oven dried meat had the highest ash, F. extract, C. protein and C. fat content compared with other processing methods. In the same vein, high potassium, iron and magnesium content was recorded in oven dry cane rat. Furthermore, there are presence of Staphylococcus aureus, Bacillus spp, Salmonella spp and Escherichia coli in the meat samples which can cause serious health problems. To encourage sustainable utilization of bush meat consumption in urban and rural areas, oven dried bush meat is the most appropriate method of processing, this should be used and encourage by the seller.