scholarly journals Comparison of Four Different Culture Media for Isolation and Growth of Type II and Type I/III Mycobacterium avium subsp. paratuberculosis Strains Isolated from Cattle and Goats

2006 ◽  
Vol 72 (9) ◽  
pp. 5927-5932 ◽  
Author(s):  
Luc�a de Juan ◽  
Julio �lvarez ◽  
Beatriz Romero ◽  
Javier Bezos ◽  
Elena Castellanos ◽  
...  
Keyword(s):  
Incubation Period ◽  
Mycobacterium Avium ◽  
Culture Media ◽  
Egg Yolk ◽  
Disease Diagnosis ◽  
Type I ◽  
Type Ii ◽  
Sodium Pyruvate ◽  
Tissue Samples ◽  
Solid Media

ABSTRACT Culture is considered the definitive technique for Johne's disease diagnosis, and it is essential for later applications of certain molecular typing techniques. In this study, we have tested four solid media (Herrold's egg yolk medium [HEYM] with sodium pyruvate and mycobactin [HEYMm-SP], HEYM with mycobactin and without sodium pyruvate [HEYMm], Middlebrook 7H11 with mycobactin [Mm], and L�wenstein-Jensen with mycobactin [LJm]) for isolation of Mycobacterium avium subsp. paratuberculosis strains in 319 tissue samples from cattle herds and goat flocks. We have shown that each of the two main groups of M. avium subsp. paratuberculosis (type II and type I/III) has different requirements for growth in the culture media studied. The recommended solid media for isolation of type I/III strains are LJm and Mm, since the combination of both media allowed the recovery of all these strains. The most widespread culture medium, HEYM, is not suitable for the isolation of this group of M. avium subsp. paratuberculosis strains. Regarding the type II strains, HEYMm-SP was the medium where more strains were isolated, but the other three media are also needed in order to recover all type II strains. The incubation period is also related to the strain type. In conclusion, because the type of strain cannot be known in advance of culture, coupled with the fact that cattle and goats can be infected with both groups of strains, we recommend the use of the four solid media and the prolongation of the incubation period to more than 6 months to detect paratuberculous herds/flocks and to determine the true prevalence of the infection.

scholarly journals Comparison of four culture media for isolation of Mycobacterium avium complex from porcine tissues

1979 ◽  
Vol 9 (2) ◽  
pp. 194-196
Author(s):  
C O Thoen ◽  
E M Himes ◽  
J L Jarnagin ◽  
R Harrington
Keyword(s):  
Mycobacterium Avium ◽  
Mycobacterium Avium Complex ◽  
Culture Media ◽  
Agar Medium ◽  
Egg Yolk ◽  
Sodium Pyruvate ◽  
Acid Fast Bacilli ◽  
Lowenstein Jensen ◽  
Time Required ◽  
Porcine Tissues

The efficiency of four culture media was compared for the isolation of Mycobacterium avium complex from 197 procine tissues. In 82 tissues with microscopic granulomas and acid-fast bacilli, a significantly greater number of isolates were obtained on Middlebrook 7H10 medium with sodium pyruvate than on Stonebrink medium, Herrold egg yolk agar medium, or Lowenstein-Jensen medium (P=0.01). In 46 tissues in which no microscopic granulomas or acid-fast bacilli were observed, a significantly greater number of isolates were made on Middlebrook 7H10 medium or Herrold egg yolk agar medium than on Stonebrink medium or on Lowenstein-Jensen medium (P=0.01). The time required to grow M. avium complex on Lowenstein-Jensen medium was significantly greater than the time required to observe growth on Stonebrink, Middlebrook 7H10, or Herrold egg yolk agar medium (p=0.001).

scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

Muscle Biopsy Samples for Histochemical Processing: Alterations Induced by Storage

1988 ◽  
Vol 25 (1) ◽  
pp. 77-82 ◽  
Author(s):  
K. G. Braund ◽  
K. A. Amling
Keyword(s):  
Skeletal Muscle ◽  
Cell Morphology ◽  
Type I ◽  
Type Ii ◽  
Tissue Samples ◽  
Frozen Tissue ◽  
Fresh Frozen ◽  
Healthy Dogs ◽  
Morphology And Morphometry ◽  
Type Ii Fibers

Skeletal muscle samples from two healthy dogs were stored in ice at 0 C for up to 30 hours to examine the influence of time on cell morphology and morphometry. Cytochemical and histochemical properties of muscle to 18 hours were not markedly different from fresh frozen tissue. Samples stored to 30 hours were still satisfactory, despite a decline and unevenness in depth of staining. Morphometry from samples stored at 0 C for 6 hours or longer is not recommended, due to the statistically significant increase in diameter (from 21 to 25%) of type I and type II fibers.

Evidence of the three main clonalToxoplasma gondiilineages from wild mammalian carnivores in the UK

Parasitology ◽  
2013 ◽  
Vol 140 (14) ◽  
pp. 1768-1776 ◽  
Author(s):  
A. BURRELLS ◽  
P. M. BARTLEY ◽  
I. A. ZIMMER ◽  
S. ROY ◽  
A. C. KITCHENER ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Rflp Markers ◽  
Type I ◽  
Type Ii ◽  
Red Foxes ◽  
Mammal Species ◽  
Tissue Samples ◽  
Pcr Rflp ◽  
Few Data ◽  
The Uk

SUMMARYToxoplasma gondiiis a zoonotic pathogen defined by three main clonal lineages (types I, II, III), of which type II is most common in Europe. Very few data exist on the prevalence and genotypes ofT. gondiiin the UK. Wildlife can act as sentinel species forT. gondiigenotypes present in the environment, which may subsequently be transmitted to livestock and humans. DNA was extracted from tissue samples of wild British carnivores, including 99 ferrets, 83 red foxes, 70 polecats, 65 mink, 64 badgers and 9 stoats. Parasite DNA was detected using a nested ITS1 PCR specific forT. gondii, PCR positive samples were subsequently genotyped using five PCR–RFLP markers.Toxoplasma gondiiDNA was detected within all these mammal species and prevalence varied from 6·0 to 44·4% depending on the host. PCR–RFLP genotyping identified type II as the predominant lineage, but type III and type I alleles were also identified. No atypical or mixed genotypes were identified within these animals. This study demonstrates the presence of alleles for all three clonal lineages with potential for transmission to cats and livestock. This is the first DNA-based study ofT. gondiiprevalence and genotypes across a broad range of wild British carnivores.

scholarly journals Comparative histochemical and morphometric analysis of muscle fibers of the psoas muscle in individuals of both genders with ageing

2020 ◽  
pp. 107-107
Author(s):  
Vladimir Antic ◽  
Nenad Stojiljkovic ◽  
Milorad Antic
Keyword(s):  
Morphometric Analysis ◽  
Statistical Significance ◽  
Muscle Fibers ◽  
Age Groups ◽  
Psoas Muscle ◽  
Iliopsoas Muscle ◽  
Type I ◽  
Type Ii ◽  
Tissue Samples ◽  
The Right

Background/Aim. In this paper, we analyzed type I and type II muscle fibers of the iliopsoas muscle in persons of both genders with ageing. The aim of this study was to detect the presence and distribution of types I and II muscle fibers in the human psoas muscle using the hematoxylin and eosin method in individuals of different ages and genders. Methods. The material consisted of tissue samples of the right iliopsoas muscle taken from 30 adult cadavers (18 males and 12 females), aged from 30 to 90 years, divided into three age groups. The material was obtained from the Institute of Forensic Medicine, Faculty of Medicine University of Nis. Results. The values of astereological parameters (area, perimeter and Feret's diameter) of type I and type II muscle fibers were higher in male cases, although without any statistical significance. Conclusion. Based on the histochemical and morphometric analysis, the conclusion was drawn that after 70 years of life there occurred a loss of type II muscle fibers, more conspicuous in female cases.

scholarly journals Comparison of Methods for Processing Drinking Water Samples for the Isolation of Mycobacterium avium and Mycobacterium intracellulare

2008 ◽  
Vol 74 (10) ◽  
pp. 3094-3098 ◽  
Author(s):  
Rachel Thomson ◽  
Robyn Carter ◽  
Chris Gilpin ◽  
Chris Coulter ◽  
Megan Hargreaves
Keyword(s):  
Mycobacterium Avium ◽  
Water Samples ◽  
Culture Media ◽  
Tap Water ◽  
Cetylpyridinium Chloride ◽  
Sodium Thiosulfate ◽  
Mycobacterium Intracellulare ◽  
Solid Media ◽  
Lowenstein Jensen ◽  
Mycobacterial Growth

ABSTRACT Several protocols for isolation of mycobacteria from water exist, but there is no established standard method. This study compared methods of processing potable water samples for the isolation of Mycobacterium avium and Mycobacterium intracellulare using spiked sterilized water and tap water decontaminated using 0.005% cetylpyridinium chloride (CPC). Samples were concentrated by centrifugation or filtration and inoculated onto Middlebrook 7H10 and 7H11 plates and Lowenstein-Jensen slants and into mycobacterial growth indicator tubes with or without polymyxin, azlocillin, nalidixic acid, trimethoprim, and amphotericin B. The solid media were incubated at 32°C, at 35°C, and at 35°C with CO2 and read weekly. The results suggest that filtration of water for the isolation of mycobacteria is a more sensitive method for concentration than centrifugation. The addition of sodium thiosulfate may not be necessary and may reduce the yield. Middlebrook M7H10 and 7H11 were equally sensitive culture media. CPC decontamination, while effective for reducing growth of contaminants, also significantly reduces mycobacterial numbers. There was no difference at 3 weeks between the different incubation temperatures.

Tetracyclines Inhibit Protein Glycation in Experimental Diabetes

1998 ◽  
Vol 12 (1) ◽  
pp. 152-158 ◽  
Author(s):  
M.E. Ryan ◽  
N.S. Ramamurthy ◽  
L.M. Golub
Keyword(s):  
Diabetic Rats ◽  
Protein Glycation ◽  
Type I ◽  
Type Ii ◽  
Tissue Samples ◽  
Glycated Proteins ◽  
Uncontrolled Diabetes ◽  
Skin Collagen ◽  
Micro Organisms

Glycation of proteins, which is accelerated in the diabetic state, has been implicated in many of the long-term complications of diabetes. This process can be inhibited by members of the tetracycline family of compounds. This novel finding is supported by studies conducted on drug (streptozotocin)induced Type I and genetic (ZDF/Gmi- fa/fa) Type II diabetic rats. These animals were orally gavaged daily with 5 mg of doxycycline and a variety of non-antimicrobial chemically modified tetracycline derivatives for time periods of 3 weeks to 11 months, while control untreated diabetic and nondiabetic animals were gavaged with vehicle alone (2% CMC). Blood and tissue samples were collected and analyzed for glucose and glycated proteins. None of the treatments had any effect on the severity of hyperglycemia or the intracellular glycation of hemoglobin of either Type I or II diabetic animals. However, the tetracycline analogues did affect the extracellular glycation of several proteins such as those found in the serum as well as skin collagen. In the Type II (ZDF) animals, initial mortality (3-5 months) was seen only in the doxycycline-treated animals, associated with infection by tetracycline-resistant micro-organisms, which was eventually surpassed by mortality rates in the untreated diabetics (6-9 months). CMT treatment not only decreased mortality but also increased longevity in the Type II diabetic animals, most likely by preventing the development of a number of long-term complications of uncontrolled diabetes, including glycation of proteins, that eventually lead to the demise of untreated diabetic animals.

CALLUS INDUCTION AND PLANT REGENERATION FROM IMMATURE EMBRYOS CULTURE OF TROPICAL MAIZE

2018 ◽  
Vol 17 (3) ◽  
pp. 359
Author(s):  
MARIA JOSÉ VILAÇA VASCONCELOS ◽  
MAURICIO SCHUSTERSCHITZ ANTUNES ◽  
MAURÍLIO FERNANDES DE OLIVEIRA ◽  
MAURÍCIO ANTÔNIO LOPES ◽  
JOSÉ EDSON FONTES FIGUEIREDO
Keyword(s):  
Tissue Culture ◽  
Zea Mays ◽  
Callus Induction ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Immature Embryos ◽  
Type I ◽  
Type Ii ◽  
Tropical Maize

ABSTRACT - The development of protocols to overcome the current limitations of callus induction and in vitro regeneration of highly recalcitrant tropical maize is crucial for plant genetic transformation. The ability of embryogenic callus (EC) formation of 46 tropical maize hybrids and 14 inbred lines was tested using N6 medium with the following changes: medium A (N6 + 15 μM dicamba + 25 mM L-proline + 88 mM AgNO3), medium B (N6 + 30 μM dicamba + 25 mM L-proline + 88 mM AgNO3), medium C (N6+ 30 μM dicamba + 6 mM L-proline), and medium D (N6+ 30 μM dicamba + 25 mM L-proline). Compact (Type I) and friable (Type II) callus were induced in the four media. Twenty genotypes produced callus in all media, and four genotypes (CO32, AG8012, CMS477BC4F2, and CMS-HGZ10) produced the highest number of callus (114, 134, 131, and 126 calli, respectively). All immature embryos of ten genotypes produced callus in at least one medium, while eight genotypes were highly recalcitrant, and they did not produce any callus. The frequencies of EC ranged from 0% to 38%, and the highest rate of EC was observed on medium B (0.40) with a total of 865 calli, and the lowest induction rate was obtained with medium D (0.29) with 555 calli (P= 0.05). From the seventy-two EC of 26 genotypes transferred to Murashige & Skoog regeneration medium, twenty-four (66.7%) differentiated into green plants which produced seeds in R0 and R1 generations, and twelve (33.3%) developed into albino plants. The results demonstrated that the problem of the recalcitrant genotypes can be, at least partially, overcome by using immature embryos as explants together with tissue culture media formulations.Keywords: Zea mays, immature embryos, somatic embryogenesis, tissue culture, type I and type II calli. INDUÇÃO DE CALO E REGENERAÇÃO DE PLANTAS DA CULTURA DE EMBRIÕES IMATUROS DE MILHO TROPICAL  RESUMO - O desenvolvimento de protocolos para superar as limitações atuais de indução de calos e regeneração in vitro de milho tropical altamente recalcitrante é crucial para a transformação genética de plantas. Foi testada a capacidade de formação de calos embriogênicos (CE) de 46 híbridos de milho tropicais e 14 linhagens cultivadas em Meio N6 com as seguintes alterações: Meio A (N6 + 15 μM dicamba + 25 mM L-prolina + 88 mM AgNO3), Meio B (N6 + 30 μM de dicamba + 25 mM de L-prolina + 88 μM de AgNO3), Meio C (N6 + 30 μM de dicamba + 6 mM de L-prolina) e Meio D (N6 + 30 μM de dicamba + 25 mM de L-prolina). Calos compactos (tipo I) e friáveis (tipo II) foram induzidos nos quatro meios de cultura. Vinte genótipos produziram calos em todos os meios, e quatro genótipos (CO32, AG8012, CMS477BC4F2 e CMS-HGZ10) produziram o maior número de calos (114, 134, 131 e 126 calos, respectivamente). Todos os embriões imaturos de dez genótipos produziram calos em pelo menos um meio, enquanto oito genótipos foram altamente recalcitrantes e não produziram nenhum calo. As frequências de CE variaram de 0% a 38%, e a maior taxa de CE foi observada no meio B (0,40), com um total de 865 calos, e a menor taxa de indução com o meio D (0,29), com 555 calos (P = 0,05). Dos setenta e dois CE de 26 genótipos transferidos para meio de regeneração Murashige & Skoog, vinte e quatro (66,7%) se diferenciaram em plantas verdes que produziram sementes nas gerações R0 e R1, e doze (33,3%) se desenvolveram em plantas albinas. O problema relacionado dos genótipos recalcitrantes de milho pode ser, pelo menos parcialmente, contornado com o uso de explantes de embriões imaturos juntamente com a formulação dos meios de cultura.Palavras-chave: Zea mays, embriões imaturos, embriogênese somática, cultura de tecidos, calos tipo I e tipo II.

scholarly journals Discovery of Stable and Variable Differences in the Mycobacterium avium subsp. paratuberculosis Type I, II, and III Genomes by Pan-Genome Microarray Analysis

2008 ◽  
Vol 75 (3) ◽  
pp. 676-686 ◽  
Author(s):  
Elena Castellanos ◽  
Alicia Aranaz ◽  
Katherine A. Gould ◽  
Richard Linedale ◽  
Karen Stevenson ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Colony Growth ◽  
Cell Entry ◽  
Genomic Islands ◽  
Type I ◽  
Type Ii ◽  
Test Panel ◽  
Repeat Sequences ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Genomic Regions

ABSTRACT Mycobacterium avium subsp. paratuberculosis is an important animal pathogen widely disseminated in the environment that has also been associated with Crohn's disease in humans. Three M. avium subsp. paratuberculosis genomotypes are recognized, but genomic differences have not been fully described. To further investigate these potential differences, a 60-mer oligonucleotide microarray (designated the MAPAC array), based on the combined genomes of M. avium subsp. paratuberculosis (strain K-10) and Mycobacterium avium subsp. hominissuis (strain 104), was designed and validated. By use of a test panel of defined M. avium subsp. paratuberculosis strains, the MAPAC array was able to identify a set of large sequence polymorphisms (LSPs) diagnostic for each of the three major M. avium subsp. paratuberculosis types. M. avium subsp. paratuberculosis type II strains contained a smaller genomic complement than M. avium subsp. paratuberculosis type I and M. avium subsp. paratuberculosis type III genomotypes, which included a set of genomic regions also found in M. avium subsp. hominissuis 104. Specific PCRs for genes within LSPs that differentiated M. avium subsp. paratuberculosis types were devised and shown to accurately screen a panel (n = 78) of M. avium subsp. paratuberculosis strains. Analysis of insertion/deletion region INDEL12 showed deletion events causing a reduction in the complement of mycobacterial cell entry genes in M. avium subsp. paratuberculosis type II strains and significantly altering the coding of a major immunologic protein (MPT64) associated with persistence and granuloma formation. Analysis of MAPAC data also identified signal variations in several genomic regions, termed variable genomic islands (vGIs), suggestive of transient duplication/deletion events. vGIs contained significantly low GC% and were immediately flanked by insertion sequences, integrases, or short inverted repeat sequences. Quantitative PCR demonstrated that variation in vGI signals could be associated with colony growth rate and morphology.

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