Introduction
Avibacterium paragallinarum
Avibacterium paragallinarum, previously known as Haemophilus paragallinarum, is a pathogenic bacteria that cause infectious coryza, an acute respiratory diseases that associate with substantial losses in poultry industry worldwide (Blackall et al., 2005). Early descriptions believed the pathogen of infectious coryza was Haemophilus gallinarum, which requires hemin and NADH as compulsory growth factors. In 1960s, studies on bacteria isolates recovered from the infectious coryza cases showed that the growth of all isolates were only NADH-dependent, leading to the identification of the new species, Haemophilus paragallinarum (Blackall & Soriano, 2008). In recent years, NADH-independent H. paragallinarum strains have been reported from South Africa, Mexico and Peru (Blackall, 1999; Soriano-Vargas et al., 2013; Falconi-Agapito et al., 2015), revealing the existence of geographic influence on bacteria distribution.
H. paragallinarum belongs to the Pasteurellaceae family. Like some of the pathogens for avian diseases, it has only been identified with chicken host. With the host-specificity character, the Avibacterium genus was established within Pasteurellaceae family in 2005, including Avibacterium gallinarum, Avibacterium paragallinarum, Avibacterium avium and Avibacterium volantium. 16S ribosomal RNA analysis revealed that at least 96.8% sequence similarity was identified within this genus. This genus was also separable from all taxa
See Table 1 and Flow Chart 1 for results of Bacteria # 1 and Table 2 and Flow Chart 2 for results of Bacteria # 2.
Often scientists work with bacteria that do not come in a labeled test tube— for example, bacterial samples taken from infected human tissue or from the soil—and the scientist must then identify the unknown microorganism in order to understand what behavior to expect from the organism, for example, a certain type of infection or antibiotic resistance. However, because of the relatively few forms of bacteria compared to animals and because of the lack of bacterial fossil records due to their asexually reproductive nature, the taxonomy used to classify animals cannot be applied to bacteria (Brown 275). In order to classify unknown bacteria, a variety of physiological and metabolic tests are available to narrow a sample down from the fathomless number of possibilities into a more manageable range. Once these tests have been performed, the researcher can consult Bergey’s Manual of Determinative Bacteriology, a systematically arranged and continually updated collection of all known bacteria based on their structure, metabolism, and other attributes.
My unknown organism #6 is Morganella morganii, which is a gram-negative bacillus rods commonly found in the environment and also in the intestinal tracts of humans, mammals, and reptiles as a normal flora. (3, 5) This bacterium Morganella morganii, was first discovered in the 1906 by a British bacteriologist named H. de R. Morgan. (2) Despite its wide distribution, it is an uncommon cause of community-acquired infection and is most often encountered inpostoperative and other nosocomial settings. (2, 3) Morganella morganii infections respond well to appropriate antibiotic therapy; however, its
According to the “Microbiology Laboratory Manual,” lab 27 focuses on the family Enterobacteriaceae, this is one of the largest families of bacteria (Wong et al., n.d.). Also all of the members are gram-negative, anaerobe straight rods and can commonly be located in mammal’s intestines (Wong et al., n.d.). All the members Enterobacter aerogenes throughout this report will be referred to as, E. aerogenes. There were several examples where E. aerogenes was tested or used, the first example is a One-pot synthesis, the next example is where it is used for dark fermentative biohydrogen production, and the last example is where it is analyzed in groundwater in North Central Nigeria.
Bacteria are ubiquitous; they can be found on the skin, in the soil, and inside the body. Because of the very nature of this ubiquity, it is important to be able to determine between different strains of bacteria. An example of this is determining the causative agent for a disease so that the patient will be treated with the appropriate antibiotics. It may be important to determine the bacteria in a certain region, because like with enteric bacteria, it is normal to find them in the digestive tract as they are in a symbiotic relationship with our bodies in this area; however, they also cause opportunistic infections in places outside of the digestive tract to our detriment, such as with a urinary tract infection. Some strains of bacteria are common to nosocomial infections, and identifying these bacteria as such helps create the guidelines for healthcare workers in antiseptic technique. All of the morphology and characteristics of each strain of bacteria help us to better understand the role of bacteria in the body as well as helps us understand how they can cause illness, and what treatment regimen to set in place. In lab this semester, a sample of unknown
Preliminary studies help identify Genus species of bacteria. Two different preliminary study pathways must be used since two different pathogens were found in the sample. A dilution and a quadrant streak are the ideal methods to separate pure cultures of bacteria. MacConkey Agar and CAN (MAC) is a selective media that is used for the cultivation of gram negative bacteria. (PEA) is a selective media that is used
I inoculated a T-Soy agar with bacteria number 118, for this I used a streak isolation method. Next, in order to distinguish between Gram positive and Gram negative I used a streak isolation technique on a CNA plate, then performed the same exact procedure on a MacConkey plate. The results from the CNA plate showed the Gram Positive bacteria was an Alpha hemolyzer. Next, I used a P Disc on a T-Soy agar inoculated with bacteria 118 and determined the Gram Positive bacteria was not sensitive to P Disc antibiotics. This revealed the Gram Positive bacteria to be Streptococcus Mitis. The results from the MacConkey plate proved the Gram Negative bacteria to be a lactose fermenter. With the Gram Negative bacteria I performed a lysine test with positive results. Next, I performed an ornithine test on the Gram Negative bacteria, with negative results, therefore I concluded the Gram Negative bacteria was Klebsiella pneumoniae.
PCR is the amplification of DNA by denaturing, annealing, and extension of a DNA template. Specific sequences can be amplified using single-stranded DNA that complements the target sequences known as primers. This process heats DNA until the strands separate, then primers bind to the target regions. DNA polymerase enzymes and single base nucleotides (dNTPs) are used to synthesize new strands of DNA to the target sequence. The end product will contain large quantities of the target sequence (Bean et al. 2015). The most notable in phylogenic studies is the 16S rRNA gene, because of it’s highly conserved primer-binding site and hyper variable regions that provide species-specific sequences within bacteria and archaea (Kolbert and Persin 1999). This gene is a component of the 30S small subunit of prokaryotic ribosome’s and serves as the primary site of protein synthesis. (Woese and Fox 1977). The 16S rRNA sequence can be amplified and matched to national databases provided by the National Center for Biotechnology Information (NCBI) using software termed Basic Local Alignment Search Tool (BLAST) to find regions of similarity between biological sequences for bacterial identification. Thus, providing a cost effective and timely method when compared to biochemical
The most represented classes were Acidobacteria and Alphaproteobacteria, which together comprised almost 30% of all bacterial amplicons, followed by the considerably lower representation of Deltaproteobacteria, DA052, and Sphingobacteria (Table 1S). Members of the dominant classes Actinobacteria, Alphaproteobacteria, and Sphingobacteria showed similar patterns between sample sites. However, Deltaproteobacteria had a higher relative abundance at RP in comparison with SB or HH1, and Elusimicrobia was in higher abundance at RP than at HH1. Levels of Nitrospirae and Spirochaetes were the highest at RP than in all other samples.
The purpose of this project was to identify the identities of two unknown bacteria in a mixed broth culture by using several separation methods. To separate the organisms, a four-way streak plate technique was used to isolate the two unknown bacteria into separate visible colonies. Then after each colony were clearly isolated; the two unknowns were processed through Gram staining test to determine the Gram stain and morphology. After Gram staining, a carbohydrate test was performed on each unknown to determine if it had glucose, sucrose, or lactose fermentation. The results of the sugar test help determining which biochemical test should be performed next. The Gram positive organism was tested through a carbohydrate fermentation test, then further tested to confirm its identity through an indole and catalase test. The Gram negative organism was tested through carbohydrate fermentation test, then further tested to confirmed its identity through an indole, and TSIA test. After running four biochemical tests, the results conclude that the Gram positive unknown was Staphylococcus aureus. S. aureus was identified based on the fermentation results of the glucose test, negative indole test, and a positive catalase production. S. aureus is a Gram positive circular shaped bacterium that is very common in the U.S and is normally found in the nose, respiratory tract, and on the skin. This bacterium is usually the most common cause of infections after injury or surgery.
The bases of this experiment was to discover the identify of the unknown from three possible specimens: Klebsiella pneumonia, Escherichia coli, and Enterobacter aerogenes. Utilizaing the T streak technique, the bacteria was isolated into pure colonies for further study. The Gram Stain method was used to identity the morhphology of the bacteria such as the shape and whether the bacteria was Gram positive or Gram negative. Biochemical test were also used to help identify the unknown bacteria. The biochemical test used was the Triple Sugar Iron Agar, Sulfur Indole Motility test, Methyl Red test, Voges-Proskauer test, Citrate test, Urease test, and the Gelatin test. After observing the morphology of the bacteria using the Gram Stain method and utilizing all the possible biochemical test, the bacteria was identified to be Enterobacter aerogenes.
pneumoniae I believe I have a subspeciesof K. pneumoniae perhaps K. pheumoniae ozaenae or K. pneumoniae rhinoscleromatis. Indole, MR-VP and Citrate will vary among different subspecies according to bergey's manual of determinative bacteriology. The reason why I decided on K. pneumoniae over E. aerogenes is because of the gelatin and motile test. According to bergey's manual, Enterobacter liquefies gelatin very slowly and in addition E. aerogenes will test positive for motility. To further differentiate K. pneumoniae from Enterobacter and narrow down the exact species of K. pneumoniae the urea hydrolysis assay could have been the determining factor. Subspecies ozaenae and rhinosclermatis will result in a negative reaction for VP but bergey’s Manual indicates a positive reaction for
A highly conserved gene will be used to identify a prokaryotic species isolated from the body. Fundamental lab techniques will be also explored and utilized, such as amplifying using PCR, cloning, and transforming the gene into a host cell. DNA electrophoresis and specific substrate plating will serve as analysis check points. The final product will be sequenced and compared to similar species to observe phylogenetic relationships.
The bacteria that was contained within Unknown tube #12 is believed to be Pseudomonas aeruginosa, Figure 1. The bacteria tested to be Gram Stain negative, producing a pink, red color retained from the staining process. When the species of bacteria was plated on nutrient media, the cells produced an irregular and spreading configuration as shown in Figure 2. This same plating test provided the margins and elevation, lobate and hilly, respectively. The specimen was stabbed in a Fluid Thioglycollate Medium (FTM) tube using an inoculated loop of the bacteria. The results of this experimentation indicate the type of oxygen requirement of the bacteria. The test found the bacteria to be aerobic as colonies of the bacteria began to form along the top of the FTM tube (Manual 2017).
The identification of an unknown sample, particularly in the environment, can be the matter of life or death. Identifying an unknown environmental sample has countless of opportunities and benefits. Identifying pathogens in the environment can save the lives of thousands or even millions of people. Some bacteria are opportunistic pathogens such as Pseudomonas. These pathogens do not commonly infect healthy people, but rather take any opportunity possible such as an open wound or an immunocompromised system. Knowing the bacteria which can prevent or promote the growth of agriculture can help to feed populations. Being able to identify bacteria is a task not to be taken lightly. Many different tests are possible to determine a specific bacterium. Often it takes a combination of several different tests to narrow down and identify a specific genus and species.