VP Test Research Articles

Isolation and Characterisation of Pink Pigmented Facultative Methylotrophs (PPFM) from Malaysian Ulam Leaves

This study aims to isolate and characterise the Pink Pigmented Facultative Methylotrophs (PPFM) bacteria from the leaves of common Malaysian table salad (ulam). The colonies of PPFM bacteria were obtained using selectively modified Pseudomonas agar based on the appearance of the pink pigment colony. The three selected isolates labelled OJ4, OJ154 [Oenanthe javanica (Blume) DC.] and ML8 [from Melicope lunu-ankenda (Gaertn.) T. G. Hartley] were chosen for characterisation. The result showed the PPFM bacteria colonies (log CFU/g) at the leaf surface of C. caudatus are 4.4 ± 0.1, significantly higher than O. javanica, 3.8 ± 0.2, and M. lunu-ankenda, 3.2 ± 0.1. The selected isolates belong to the Gram-negative group, motile with rod shape with size [length (l) × width (w)] in µm unit 4.3 ± 1.1 × 1.6 ± 0.6, 5.4 ± 0.2 × 1.2 ± 0.1, and 3.5 ± 0.7 × 1.0 ± 0.0, respectively. They show positive urease, catalase, and oxidase activities, while none of them can degrade starch, gelatine, or cellulose, as well as glucose fermentation (MR test) and metabolism actions (VP test), producing indole and hydrogen sulphide gaseous. Only isolate OJ154 demonstrates positive casein hydrolysis and nitrate reduction activities, while only isolate ML8 can utilise citrate but not in lipid degradation. Their sequence analysis of the 16S rRNA indicated that OJ4 and ML8 are Methylobacterium radiotolerans with a similarity of 99%, whereas, OJ154 is Methylorubrum salsuginis with a similarity of 99%. To conclude, PPFM bacteria from the leaves of C. caudatus, O. javanica, and M. lunu-ankenda have been isolated and characterised in particular.

Biochemical characterization of bacteria isolated from Rhizosphere soils of sugarcane grown in Uttar Pradesh

Rhizospheric soil samples of Sugarcane fields were collected from distinct regions of U.P. Further, these soil samples were used for the isolation and characterization of bacteria to identify their maximum PGPR traits and biochemical potential. A total of ten bacterial isolates were collected from the soil samples and sub-cultured in the nutrient agar plates and characterized through Biochemical test. One bacterial strain was found negative and the other 9 isolates were found positive for gram staining. All bacterial isolates were positive for citrate test. 3 isolates were found MR positive and 2 positives for VP test. 5 isolates were positive for gelatinase test and 9 isolates were positive for nitrate reductase test. 2 isolates were positive for starch hydrolysis test, while 7 isolates were found positive for phosphate solubilization test and 3 isolates were positive for siderophore production test. These results showed that the bacterial isolates from soil sample were Azotobacter Species, Azospirillum Species, Pseudomonas species, Bacillus Species, Rhizobium Species, Serratia Species and E. coli. These bacterial strains followed the maximum PGPR traits. After proper validation, these bacterial strains could be used for the bio-fertilizer formation for crop production enhancement by improving soil fertility.

Determination of the Anti-bacterial potency of Allium sativum (Garlic) extract against Agrobacterium tumefaciens causing Crown Gall Disease

The isolated crown galls (a bacterial plant disease) from Pongamia pinnata leaf were investigated to identify the causative agent. The SEM analysis revealed that the causative agent in crown gall appeared to be a slender, pink and rod-shaped bacterium. The morphological, cultural, biochemical and tumorogenesis analyses were performed to identify causative organisms in the crown gall disease. The isolate was negative for Indole, MR and VP tests, but positive for Citrate, TSI, Urease, Catalase and Oxidase tests. The causative organism also developed a tumor-like sport during the potato disc assay after 3rd week of incubation. The causative agent in crown gall was identified as Agrobacterium tumefaciens. The ethanolic extract of Allium sativum produced an inhibitory zone of 15 mm against this isolate. The current study proved that ethanolic extract of garlic can be effectively used to treat crown gall disease in the economically valuable dicot plant of Pongamia pinnata.

Isolation, Identification and Risk Factors of Staphylococcus aureus Bacteria in Dairy Goat Milk with Subclinical Mastitis in Siliragung District, Banyuwangi Regency

Mastitis is an inflammation of the mammary glands that can cause a sharp reduction in milk production and income for farmers. Subclinical mastitis is mastitis without clinical symptoms but can result in a decrease in the quantity and quality of milk production. One of the main pathogens that cause mastitis namely Staphyloccocus aureus. S. aureus become the biggest problem of mastitis because the cure rate is classified as very low. The risk factors for the occurrence of mastitis, both clinical and subclinical mastitis, can cause a decrease in milk production caused by several factors, including genetic, feed and environmental factors. This study aims to determine the presence of subclinical mastitis in milk of dairy goats with sufferers in Siliragung District, Banyuwangi Regency. Sample testing was carried out at the SIKIA Instrument Laboratory, Airlangga University. Isolation was carried out using MSA (Mannitol Salt Agar) media for further identification consisting of Gram staining and biochemical tests. Some of the biochemical tests used are the catalase test, the coagulase test and the VP test. The results of isolation, observation of characteristics and identification of bacteria in the research that has been carried out show that 64.51% (40/62) of the samples examined were positively identified S. aureus. Results showed that the risk factors for causing subclinical mastitis included cleaning the cage, the number of lactating goats, washing hands before milking, manure and waste management, weaning age and lactation age.

Isolation and characterization of Streptococcus suis from pig farms in Sichuan

In this experiment, bacteria were isolated by aseptically collecting tissue samples of heart, spleen, liver, lymph node, and joint fluid from diseased and dead pigs, and were identified by colony morphology observation, Gram staining microscopy, biochemical experiments, PCR identification, pathogenicity test, and drug sensitivity test. The results showed that the isolated strains were Gram-positive, long-chained, or bipartite coccoid bacteria. It could ferment most sugars, and the VP test and indigo substrate test were negative, consistent with the physiological characteristics of Streptococcus suis. The PCR method was used to identify the serotypes, and about 500 bp of podocarp serotype 9-specific bands could be amplified. Pathogenicity test showed that mice became depressed 6h after intraperitoneal inoculation with the isolate, and died within 24h after rapid breathing, slow movement, and loss of appetite. The results of the drug sensitivity test showed that the isolate was highly sensitive to cefthiophene, florfenicol, cefadroxil, ciprofloxacin, ampicillin, amoxicillin, highly sensitive to sulfisoxazole, polymyxin B, enrofloxacin, doxycycline, and showed resistance to lincomycin, kanamycin, neomycin, gentamicin.

Salmonella sp. Contamination Detection in Layer Chicken Eggs Traded in Traditional Market of Medan Johor District

Salmonella sp. can contaminate layer chicken eggs and cause salmonellosis. This study aims to determine whether there is contamination by Salmonella sp. in layer chicken eggs traded in traditional markets of Medan Johor District. Sampling locations were determined by purposive sampling, including Kwala Bekala Market, Johor Market, and Tikung Market, with 90 samples. The working procedures included isolating bacteria from egg yolks using the pour plate method on SSA media, observing the bacteria morphology, calculating total Salmonella sp. colonies, Gram staining, and biochemical tests. The research results obtained indicated that the average total colonies of Salmonella sp. in Kwala Bekala Market were 1.45 x 105 CFU/g, in Johor Market they were 5.8 x 104 CFU/g, and in Tikung Market they were 3.6 x 104 CFU/g. The colony morphology of Salmonella sp. is circular and colourless, with a black spot in the center. The Salmonella sp. bacteria are Gram-negative with a red color, and are bacilli. Biochemical tests showed indole (-), MR (+), VP (-), SCA (+), and TSIA (+) test results. Based on the study results, it can be concluded that 4.44% of the positive samples were contaminated with Salmonella sp. in the traditional market of Medan Johor District and exceeded the BMCM in SNI 7388:2009.

Isolation of endophytic bacteria from bryophytes and study of their morphological, biochemical and biofilm formation properties

Aim: To isolate an endophytic bacterial community associated with bryophytes, which were collected from various locations in Mount Abu, Rajasthan, and to characterise their morphological, biochemical, and biofilm development properties. Methodology: Different bryophytes were selected from various regions of Mount Abu, Rajasthan. On an R2A agar plate, colonies with various morphologies were isolated. Biofilm development of bacterial isolates was done on a glass surface and visualised through scaning electron microscopy. Biochemical test, such as indole test, catalase test, urease test, MR and VP tests, citrate utilization test, gelatine and nitrate test, etc., were determined as per Berge’s Manual of Determinative Bacteriology for characterization of endophytic bacteria. Salkowski's technique was also used for estimating IAA production by endophytic bacteria. Results: The results confirmed that isolates belonged to two phyla groups, Firmicutes and γ-proteobacteria with predomination of Firmicutes. Scanning electron microscopy showed that among all the isolates, rod-shaped bacteria dominated. The metabolic properties of endophytic bacteria showed that most bacterial isolates metabolised glucose, maltose and sucrose, however, only few bacterial endophytes metabolized lactose. Some isolates produced IAA growth hormones, which were analysed through Salkowski's technique. Interpretation: The findings indicate that bryophytes have associated bacterial diversity. The morphological and biochemical characterization of bacteria revealed that they are members of the Firmicutes and γ-proteobacteria phyla. SEM confirmed that bacterial isolates can form biofilm on glass surfaces and produce a distinct amount of IAA in a tryptophan-supplemented medium, essential for the growth and development of bryophytes. Key words: Bryophytes, Biofilm, Endophytic bacteria

Occurrence and antimicrobial profiling of K. pneumoniae in burn patients at burn ward, Allied Hospital, Faisalabad

Background: Klebsiella pneumonia is an opportunistic pathogen causing a great array of illness both in humans and animals and responsible for high mortality rate in burn patients. Aim: To determine occurrence of K. pneumonia in burn wound patients and to check antibiotic sensitivity of K. pneumoniae. Methods: A retrospective cross sectional study was conducted and 50 samples of burn wound patients from Allied Hospital Faisalabad were taken for isolation, identification and antibiotic resistance pattern of K. pneumonia. Samples then inoculated on MacConkey agar for pure colonies. For identification gram staining and biochemical test were performed. Kirby-Bauer disk diffusion technique was used to determined antibiotic sensitivity profiling K.pneumoniae against various disks. Results: On MacConkey agar mucoid, lactose fermenting pink colonies of K. pneumoniae were observed. Among the biochemical tests, catalase, citrate and VP test were found to be positive whereas indole and methyl red tests were found to be negative. Occurrence of K. pneumoniae was 28% in burn patients. Antibiotic sensitivity profiling showed that K. pneumonia was found highly sensitive to Imipenem (88.94%), meropenem (79.32%), ertapenem (75%), moderately sensitive to aminoglycoside group (amikacin 59.13%, gentamicin 52.40%, Netilmicin 50.96%), piperacillin-tazobactam (53.84%) mildly sensitive to tobramycin (49.51%), nitrofurantoin (48.07%), levofloxacin (45.19%) Conclusion: K. pneumoniae and other gram negative bacilli are frequently associated with nosocomial burn infection. Furthermore, most isolates of K. pneumoniae from burn units were MDR K. pneumonia. Keywords: Opportunistic pathogen, Antibiotic sensitive, Burn, Mortality, Nosocomial.

Neuromuscular Paralysis is not a Barrier to Same Day Catheter Removal After HoLEP

To determine if same day catheter removal is feasible in a population of holmium laser enucleation of the prostate patients who received paralysis and if bladder pressure monitoring could predict successful catheter removal. From February 2021 to February 2022, we evaluated same day catheter removal for patients undergoing holmium laser enucleation of the prostate. Criteria for proceeding with same day catheter removal included: an uncomplicated procedure, continuous bladder irrigation weaned within 120 minutes of arrival to PACU, minimal postoperative hematuria and a bladder pressure over 30 cmH20 as measured using the VP Test device (SRS Medical). One hundred and eight patients were enrolled, with a median age of 68 years (IQR: 62.0-73.0) and prostate volume 80.8cc (IQR: 64.8-112.3). Criteria for catheter removal was met by 83/108 (76.9%) patients. Of those that attempted a trial of void, 69/83 passed for an effective pass rate of 83.1%. Median maximum detrusor pressure for those that passed and failed were 51.0 cmH20 (IQR: 40.5 -68.0) and 48.0 cmH20 (IQR: 38.8-61.8) [P=.53], respectively. Intraoperative Lasix administration was associated with a higher rate of trial of void (TOV) success and preoperative PVR was associated with lower TOV success, with OR (95%CI) of 5.8 (1.4, 29.5) [P=.02] and 0.6 (0.4, 0.9) [P=.01], respectively. Same day catheter removal is feasible in those who receive neuromuscular paralysis, with a success rate >80%. Bladder pressure monitoring did not assist with differentiating which patients will pass or fail a trial of void. Intraoperative Lasix administration may be helpful in increasing success for same day catheter removal while elevated preoperative PVR was associated with TOV failure.

Occurrence of Enterobacteriaceae in medicinal plants sold in local markets of Savar, Dhaka

Medicinal plants can be contaminated with a large diversity of potentially pathogenic bacteria. Ten medicinal plant samples Plantago psyllium, Terminalia bellirica, Terminalia chebula, Phyllanthus emblica, Ocimum basilicum, Glycyrrhiza glabra, Withania somnifera, Andrographis panicula, Bombax ceiba were collected from Amin Bazar and Savar of Savar area to study the presence and abundance of Enterobacteriaceae family bacteria in the herbal plant samples. A total of fifty five (55) bacterial isolates have been isolated from different medicinal plant samples using spread plate method on nutrient Agar media. Culture of bacterial isolate on a number of selective and differential media and biochemical tests (Catalase activity, hemolytic property, fermentation test, Starch hydrolysis, VP test, MR test, MIU test, Indole and casein test etc.) revealed the presence of different Enterobacteriaceae family bacteria in the plant samples. Out of 55 isolates 6 isolates were Salmonella sp. (11%), 2 isolates were Klebsiella sp. (3.63%) and 2 isolates were Enterobacter sp. (3.63%). Antibiotic sensitivity test of bacterial isolates showed that all the identified Enterobacteriaceae family bacteria were sensitive to Kanamycin but resistant to Amoxycilin, Aztreonam and Penicillin-G. Therefore, this study highlighted the necessity for constant quality assessment of herbal medicines to facilitate and ensure safe production and supply of medicinal products suitable for human consumption.
 Jahangirnagar University J. Biol. Sci. 10(1 & 2): 1-8, 2021 (June & December)

Characterization and Test of Endophytic Bacterial Isolates to Control Leaf Blight on Acacia Seeds (Acacia crassicarpa)

ABSTRACT Acacia is a type of plant that recommended for development of industrial forest plantations (HTI) especially pulp and paper. According to Central Statistics Agency (2020), area of ​​​​acacia HTI in Riau Province 2019 was 2,335,450 ha with a log output of 31,509,228m3 and total pulp export of 2,952.4 million US dollars. The need for paper pulp has increased every year. Problems in acacia nursery area is presence of disease caused by Xanthomonas sp. Bacterial leaf blight causes significant losses in procurement of acacia seedlings, so we need an alternative control with environmentally friendly and sustainable by utilizing potential of biological agents in controlling leaf blight. This aimed to study potential of endophytic bacterial isolates against bacterial pathogen Xanthomonas sp. by adopting a Completely Randomized Design (CRD). The results showed that there were 9 isolates of endophytic bacteria that had inhibitory power against Xanthomonas sp., bacterial isolates in hypersensitivity test did not show symptoms of necrosis in tobacco leaves, results of physiological and biochemical tests on levan test and VP test showed differences in reaction results on tested endophytic bacteria isolates and it was found isolate of endophytic bacteria B8 was effective as an antagonist agent in controlling disease. Keywords: acacia; characterization; endophytic bacteria; in vitro; in vivo; Xanthomonas sp.

Correlation of P-wave velocity with mechanical and physical properties of limestone with statistical analysis

This study aims to investigate the correlation between the P-wave velocity (Vp) and the mechanical and the physical properties of the limestone; Vp tests were conducted on over 320 limestone samples. Moreover, the effects of the mineralogical, textural, and chemical composition of limestone were also studied through thin sections, scanning electron microscopy (SEM), X-ray diffraction (XRD), and X-ray fluorescence (XRF). The relationships between the Vp and the uniaxial compressive strength (UCS), point load index (PLI(Is(50)), 2nd cycle of slake durability index (Id2), natural unit weight (γn), specific gravity (Gs(c)), water absorption by weight (WA), and porosity (n) were estimated using representative empirical equations. The empirical equations were validated by Student’s t test that has indicated the existence of strong relationships between the mechanical and physical properties of the intact limestone with Vp; the calculated t-values were higher than the t-critical value. Furthermore, the results of previously available studies were compared with the results of this study in terms of the generated equations for Vp values and the slope of a 1:1 line, which was used to appraise the predicted and measured values. This study demonstrates that the UCS, PLI(Is(50)), Id2, γn, Gs(c), WA, and n values of an intact limestone can be predicted by using Vp, which is fast, easy, economical and nondestructive test.

Isolation of Pseudomonas Species from Rhizospheric Soil and its Antagonistic Effect on Plant Pathogen

The pathogen attack on plants (such as tubers and small plants) has substantially progressed by 60% irrespective of even when the plants have strong immune system for the past five decades. It drastically affects the plant growth, yield and production. Certain pesticides and fertilizers have been tried to control the progressing pathogenic microorganisms. Although these measures have proved worthless due to the resistance shown by those pathogens and resulted in environmental pollution over a period of time. Bio control measures are well appreciated as it is ecofriendly. Antagonist microorganism (bacteria, fungi) will be used to suppress the growth of the invading pathogens. In this study, Pseudomonas species , were screened from rhisozpheric soill from the local region of Srivilliputhur. Eleven isolates from rhizospheric soil were screened and characterized by Gram staining, catalase test, Voges-Proskauer test, and oxidase test. The morphological studies of all the screened Pseudomonas isolates were observed as Gram negative. Further, all the isolates were found to be catalase positive and negative for VP test. The isolates were subjected to antagonistic effect on plant fungal pathogen such as Rhizoctonia solani. Among eleven Pseudomonas isolates four isolates showed antagonistic effect against R.solani. In the present investigation, four isolates showed potent antagonistic effect and could be used as effective bio-control agent against plant fungal pathogen R.solani.

First report of Cedecea neteri Causing Yellow rot Disease in Pleurotus pulmonarius in China.

Pleurotus pulmonarius is a popular edible fungus and widely cultivated in many areas of China. In June 2018, yellow rot (more than 10% incidence) was found on the first crop of P. pulmonarius fruiting bodies in a mushroom factory in Nanning, Guangxi Province, China. At first, yellow water-soaked lesions appeared in the infected fruiting bodies. Lesions then spread and purulent tissues were formed. Severe rot induced production of deformed fruiting bodies and offensive odor. Internal sections of the diseased tissue (approximately 0.5 × 0.5 cm) were sterilized in 75% alcohol for 30 s, rinsed three times with sterilized and deionized water, crushed and suspended in sterilized and deionized water. The suspension was spread on the Luria-Bertani (LB) medium. After incubation at 30°C for 2 days, dominant bacterial colonies were oyster white, smooth, convex, and circular. Individual colonies were transferred two times to LB medium using the conventional streak plate techniques to obtain the pure cultures. The cells were gram-negative, short rods, motile, and no capsules or endospores were observed. Using a BoJian Gram-negative bacteria biochemical analysis kit (5 CARDS, Hopebio, Qingdao, China), data were obtained and analyzed, showing that the isolated strain belongs to the Cedecea genus (positive for β-galactosidase, citric acid, arginine, sucrose, mannitol, sorbitol, D-glucose, gelatin hydrolysis and VP test but negative for H2S, urease, oxidase, indole, rhamnose, melibiose, amygdalin, lysine, ornithine, lactose, inositol and arabinose). Amplified 16S rDNA gene sequences (1,424 bp, GenBank accession No. MT925570) of the isolate using the universal primers 27f and 1492r (Lane 1991) exhibited 99.86% identity with Cedecea neteri M006 (CP009458.1). Based on its morphological characteristics, 16S rDNA sequences, and biochemical test results, the strain was identified as C. neteri. Pathogenicity tests for this strain were performed with bacterial suspensions (approximately 1 × 108 CFU/ml) after growing for 24 h in LB medium at 30°C. Mycelia of P. pulmonarius were cultivated for 60 days in plastic bags. Then young fruiting bodies were formed after induced with low temperature stimulation to serve as a host source. The prepared bacterial suspensions were directly sprayed onto the surface of three bags of fruiting bodies; another three bags were sprayed with sterilized and deionized water as negative control. All inoculated fruiting bodies were then incubated at 20°C with 90 to 95% relative humidity. All experiments were repeated three times. After 2 days, all the fruiting bodies inoculated with the bacterial suspensions showed yellow water-soaked lesions, and the normal growth of the fruiting bodies was inhibited. An offensive odor then developed along with a severe soft rot that was similar to the disease symptoms observed under natural conditions. The fruiting bodies of negative control were growing healthily with no symptoms. Koch's postulates were fulfilled by isolating bacteria from lesions on artificially inoculated fruiting bodies that were identical to the original isolates based on morphological characteristics, 16S rDNA sequences and biochemical test results. C. neteri was formally reported as a pathogen to humans that could cause bacteremia (Farmer et al. 1982). Recently, it has also been reported causing soft rot disease on mushrooms of Pholiota nameko (Yan et al. 2018) and yellow sticky disease on mushrooms of Flammulina velutipes (Yan et al. 2019). However, to the best of our knowledge, this is the first report of C. neteri-induced yellow rot disease of P. pulmonarius in China.

Supra-Versus Submaximal Cycle Ergometer Verification of VO2max in Males and Females.

This study was designed to determine the optimal intensity for verification phase testing (VP) in healthy, young adults. Thirty one young, active participants (16 females) completed a cycle ergometer graded exercise test (GXT) VO2max test and 4 VP tests at 80, 90, 100, and 105% of the maximum wattage achieved during the GXT. GXT and VP VO2max values showed a significant test x sex interaction (p = 0.02). The males elicited significantly higher VO2max values during the GXT, 80%, and 90% when compared to the 105%, (105 vs. GXT: p = 0.05; 105% vs. 80%: p < 0.01; 105% vs. 90%: p = 0.02). There were no significant differences in VO2max across the tests in the females (p > 0.05); 80% of the males achieved their highest VP VO2max during a submaximal VP test compared to only 37.5% of the females. A secondary study conducted showed excellent reliability (ICCs > 0.90) and low variation (CVs < 3%) for the 90% VP. Our findings show that a submaximal verification phase intensity is ideal for young healthy males to elicit the highest VO2max during cycle ergometer testing. For females, a range of intensities (80–105%) produce similar VO2max values. However, the 80% VP yields an unnecessarily high time to exhaustion.

Isolation, Biochemical Characterization, Antibiogram Pattern and PCR Based Confirmation of Brucella from Cows and Buffaloes

Brucellosis is a worldwide zoonotic disease that still constitutes a major public health problem in India. In the current study, Brucella were isolated from aborted samples and typed by both molecular and conventional techniques. A total 114 aborted samples were collected from cows and buffaloes in and around Anand district. Aborted samples were cultured on the Brucella agar medium and incubated for 24–48 h. Three samples from cow were found to be positive for Brucella. All the isolates were positive for catalase, oxidase and nitrate reduction while negative for urease reaction, indole test, VP test, motility examination and production of H2S. For the detection of Brucella DNA by PCR, three different genus-specific primer pairs viz., B4/B5, JPF/JPR, and F4/R2 were used. All three Brucella isolates were positive by B4/B5 and F4/R2, while two isolates were positive for JPF/JPR. For species-level identification of Brucella isolates were subjected to AMOS PCR and Bruce-ladder PCR, and were found to be B. abortus.

Investigation of biotype, genotype and virulence associated genes inPasteurella multocida capsular type A strains from the respiratory tract of cattle and their relationship with disease cases

Pasteurella (P.) multocida, which causes respiratory disease in cattle, can also be found commensally in the upper respiratory tract. Therefore, it is epidemiologically important to determine the possible difference between pathogenic and commensal strains for the diagnosis of the disease. In this study, phenotypic and genotypic characteristics and virulence associated genes of P. multocida capsular type A strains isolated from the respiratory tract of sick and healthy cattle were investigated. Also, the relationship between the disease status of animals and the characteristics of the isolates were evaluated. Total of 51 P. multocida capsular type A strains were biotyped and genotyped by using commercial bacteria identification kits and PCR-fingerprint, respectively. Four major biotypes generated from xylose and sucrose test results were detected in 39 (76.47%) isolates. Eleven different biotypes were detected in the remaining 12 isolates (23.53%), which were separated from the 4 major biotypes by mannitol, indole, sorbitol, nitrate, VP, lactose, arabinose, salicin, and rhamnose test. Four genotypes were detected in P. multocida isolates and most of them (90.2%) had genotype I profile. Virulence associated genes, such as hgbA, exbB, exbD, tbpA, ptfA, hsf-2, tadD, nanB, nanH, pmHAS, sodA, sodC, and oma87 were detected in all isolates whereas hgbB, toxA, and hsf-1 genes were not found by PCR. PfhA gene were found in 49 isolates (96.07%) but, ompH gene could be detected only in 10 (18.86%) isolates. The findings of the study indicated that P. multocida capsular type A strains generated as xylose positive/sucrose negative (biotype II) and genotype II may be associated with respiratory diseases of cattle. However, ompH gene was observed to be associated with sucrose negative isolates, but not with disease cases. As a result, it was concluded that xylose positive/sucrose negative biotype and genotype II determined in P. multocida capsular type A strains isolated from respiratory diseases of cattle, might be important epidemiological criteria in the differentiation of commensal and pathogenic isolates.

IDENTIFIKASI BIOKIMIA DAN AKTIVITAS ANTIBAKTERI ISOLAT BAKTERI ASAM LAKTAT DARI LIMBAH SAYUR SAWI (Brassica juncea L.)

Lactic Acid Bacteria (LAB) is a group of bacteria that produce lactic acid as the main product in carbohydrate fermentation. LAB has a major role to fight against pathogenic bacteria through peptide compounds. LAB can be found in vegetables that contain carbohydrates. The research objective was to determine biochemically and determine the antibacterial activity of LAB isolates from Brassica juncea L vegetable waste. Biochemical testing methods include TSIA, Indol, MR_VP, motility, temperature resistance and salt resistance tests. While testing the activity using the disc diffution method is used by the media Mueller Hinton Agar against Basillus subtilis , Escherichia coli , Staphylococcus aureus , Staphylococcus epidermidis , Streptococcus mutans , Salmonella thypimurium , and Propionibacterium acnes . Preparation of the test sample was carried out by inoculating LAB isolate stock for 1x24 hours at 37 ° C. Biochemical identification results showed that LAB isolates were able to ferment glucose, negative in indole and VP testing, positive in MR testing was characterized by obtaining acidic pH, non-motile nature, living at 37 ° C, and resistant to several salt concentrations (5-10%). The results of testing antibacterial activity based on univariate analysis showed differences in the activity of each of the test bacteria obtained p = 0,730 (p>0,05). LSD analysis showed no significant differences between all isolates. Conclusion: isolates obtained included LAB group from the family Lactobacillaceae, genus Lactobacillus sp and the antibacterial activity was broad spectrum

Isolation and Characterization of Azotobacter from Neems Rhizosphere

Nitrogen-fixing bacteria can be isolated from rhizosphere plants, and one of the N2-fixing bacteria is Azotobacter. One of the plants that can grow well in dry and barren soil conditions is Neem (Azadirachta indica A. Juss)(Mimba). The purpose of this study was to determined any Azotobacter isolated from the rhizosphere of the neem plant. The type of research is an experimental study, tracing the diversity of Azotobacter in the rhizosphere of the neem plant. The study was conducted by taking soil samples from the rhizosphere of neem plants on the Bali Island. Morphological characterization based on colony observation in terms of shape, color, margin, and texture. The isolated cells were determined based on Gram staining and determination of the presence of spores. Isolates were characterized by various biochemical methods namely nitrate, reductase, citrate utilization, methyl red, VP test, catalase (method of slip-cover), and oxidase tests. The soil sample was taken from rhizospheres of 5 plants. The results showed that the N2-fixing bacteria isolated in the five soil samples were Azotobacter paspali and Azotobacter vinelandii (Azomonas) groups. Actinomycetes were found when isolating using specific media Azotobacter on plant 2.

Isolation and Characterization of Buprofezin Tolerant Bacteria from Rhizosfer of Paddy at Marginal Land of Banyumas Regency

Buprofezin is a pesticide used to control planthoppers. The buprofezin residue may reduce the soil fertility by decreasing the essential microbes in the soil. This study aimed to isolate and characterizes buprofezin tolerance bacteria from rhizosfer of paddy at five marginal land of Srowot, Pageralang, Gunung Tugel, Tamansari, and Sokawera village, Banyumas Regency, Central Java. The bacterial isolate screened by growing on nutrient agar containing 2 ppm of buprofezin. The growing colonies were macro-morphologically observed. The growth curve and the generation time of the dominant colonies were analyzed. The selected colonies were cultured in nutrient broth containing 0, 5, 10, and 15 ppm of buprofezin. The selected colonies were characterized by gram staining, endospore staining, and biochemical test. Thirty collected-bacterial isolates showed five dominant colonies (SR1, PA1, GT2, TS4, SW1). The selected dominant isolates, SR1 colony was tolerant to buprofezin at 5 ppm and 10 ppm and GT2 at 5 ppm of buprofezin. The SR1 and GT2 were rod-shaped, gram-positive and endospore forming bacteria, white, and medium in size. The biochemical tests showed the SR1 were motile, had catalase activity, can ferment glucose, sucrose, and mannitol without gas forming, unable to ferment lactose, hydrolyze starch, positive result for MR test, but negative for urease, VP test, simmons citrate, H2S production, oxidase, and indole. The GT2 were motile, positive result for catalase test, carbohydrate fermentation (glucose, sucrose, mannitol, lactose) with gas forming, MR/VP test, simmons citrate, oxidase, and indole, but negative for starch hydrolysis, urease, and H2S production. The SR1 and GT2 were aerobic/anaerobic facultative bacteria. SR1 and GT2 were probably Bacillus sp.