Bacillus cereus nositelj plazmida pXO1 s genom pag uzrokuje u goveda s oslabljenim imunosnim sustavom smrtonosnu septikemiju sličnu bedrenici - kratko priopćenje.

Bacillus cereus is ubiquitous in nature and while most isolates appear to be harmless, some are associated with food-borne illnesses, wound infections, endocarditis, osteomyelitis, endophthalmitis and urinary tract infections in humans. Recently, a few isolates have been identified as the causative agents of anthrax-like severe pneumonia in humans, and these isolates were found to harbor most of the B. anthracis virulence plasmid pXO1. Here we report the characterization of three clinical B. cereus isolates recovered from heart blood and spleen samples of cattle which had died with ‘anthrax like’ symptoms. Apart from the cultural characterizations, primers targeting the 16S rRNA gene of B. cereus were designed and used on these isolates. The isolates were found to harbor the pXO1 plasmid and lacked pXO2 plasmid. Further characterization of the pXO1 plasmid revealed that the isolates contained pag, lef and cya genes, which code for protective antigen, lethal factor and edema factor toxins responsible for eliciting an ‘anthrax like disease’ in cattle. The sequencing and phylogenetic analysis of partial pag gene sequences of B. cereus isolates were identical to pag gene sequences on the pXO1 of B. anthracis. In a pathogenicity test on mice, B. cereus isolates, when inoculated by the intra peritoneal route, caused mortality of the mice within 6 hours post inoculation.Bacillus cereus je posvudašnja bakterija. Većina izolata te bakterije je neškodljiva, a neki mogu uzrokovati bolesti koje se prenose hranom, infekcije rana, endokarditis, osteomijelitis, endoftalmitis i infekcije mokraćnog sustava u ljudi. Nedavno je identificirano nekoliko izolata koji uzrokuju tešku upalu pluća u čovjeka sličnu onoj kod bedrenice. Ti izolati većinom nose virulentni plazmid pXO1 vrste B. anthracis. U ovom radu određena su obilježja triju kliničkih izolata vrste B. cereus izdvojenih iz krvi sadržane u srcu i uzoraka slezene goveda uginulih pod znakovima sličnima bedrenici. Osim određivanja kulturalnih obilježja, pripremljene su i početnice za gen 16S rRNA vrste B. cereus koje su bile rabljene za identifikaciju izolata. Ustanovljeno je da izolati nose plazmid pXO1, a nedostaje im plazmid pXO2. Daljnja karakterizacija plazmida pXO1 pokazala je da izolati sadrže gene pag, lef i cya koji kodiraju za zaštitni antigen, letalni čimbenik i edemski čimbenik, toksine koji su odgovorni za pojavu bolesti u goveda slične bedrenici. Određivanje slijeda i filogenetska analiza dijela sekvencija gena pag izolata B. cereus pokazala je da su oni istovjetni sekvencijama gena pag na pXO1 bakterije B. anthracis. U testu patogenosti na miševima, izolati B. cereus prouzročili su njihovo uginuće šest sati nakon intraperitonejske inokulacije

Current Trends in Nanoporous Anodized Alumina Platforms for Biosensing Applications

Pristine aluminum (Al) has received great deal of attention on fabrication of nanoporous anodized alumina (NAA) with arrays of nanosized uniform pores with controllable pore sizes and lengths by the anodization process. There are many applications of NAA in the field of biosensors due to its numerous key factors such as ease of fabrication, high surface area, chemical stability and detection of biomolecules through bioconjugation of active molecules, its rapidness, and real-time monitoring. Herein, we reviewed the recent trends on the fabrication of NAA for high sensitive biosensor platforms like bare sensors, gold coated sensors, multilayer sensors, and microfluidic device supported sensors for the detection of various biomolecules. In addition, we have discussed the future prospectus about the improvement of NAA based biosensors for the detection of biomolecules

Development of polymerase chain reaction for detection of predominant streptococcal isolates causing subclinical bovine mastitis

208-212Bovine mastitis is the most important source of loss for the growing dairy industry. Streptococci, with special reference to Streptococcus agalactiae, S. dysgalactiae and S. uberis, are the predominant pathogens causing bovine mastitis. A rapid, sensitive and specific test for the detection of these pathogens needs to be developed. To accomplish this, initially 163 milk samples were collected from various organized and unorganized sectors in and around Bangalore, India. These milk samples were screened for subclinical mastitis by somatic cell counting (SSC) and electro conduction (EC). Of these, 131 samples selected based on SCC and EC values were subjected for isolation of the organisms. Two sets of specific primers, targeting streptococcal 16S rRNA gene were designed for detection of S. agalactiae, S. dysgalactiae and S. uberis. The results of the study showed S. agalactiae as the predominant streptococci among the generally identified streptococcal species associated with subclinical bovine mastitis in dairy cattle in and around Bangalore. </span

Cytopathic effect of PPR vaccine virus strains in Vero cells

The present study describes the cytopathic effect of two different Peste des petits ruminants (PPR) vaccine virus strains presently being used in the country, in vero cells. The cytopathic effect (CPE) was visible from 4th day post infection in Sungri vaccine virus strain where as Arasur vaccine virus strain showed CPE, 36-48 hr post infection. With both vaccine virus strains the CPE in vero cells showed initial cell rounding, aggregation and syncytial development. The generalized CPE was noticed by 6th day in Sungri and by 96 hrs post infection in Arasur strain. However complete detachment of the cell monolayer was observed in Arasur strain by 120 hr, post infection. Infected coverslip cultures stained with H &amp; E and May &amp; Grunwald&amp;#8217;s Giemsa showed cell vaculation, cytoplasmic extension and syncytia comprising of five to six nuclei. Acidophilic intracytoplasmic and intranuclear inclusion bodies were also observed. Titers, HA activity and detection by s-ELISA of both the vaccine virus strains are also compared. [Vet. World 2009; 2(3.000): 93-94

PCR based confirmation of sheeppox vaccine virus

[Vet World 2010; 3(4.000): 160-

Real-time PCR confirmation of KFD seed virus.

The RNA extracted from the mouse brain suspension containing the KFD seed virus was subjected to Real-time PCR as per the procedure described by Mourya and co-workers [13]. The PCR yielded specific amplifications indicating the presence of KFD virus in the seed stock of mouse brain suspension.</p

Sections of brain showing normal architecture of cerebrum and hippocampus in control mice (Hematoxylin and Eosin staining as per the procedure of Luna [17]).

A: Normal appearance of neurons in the cerebrum, B: Normal appearance of neurons in the hippocampus.</p

In vivo titration of KFDV (MLD₅₀) by mice inoculation test.

Each dilution of the KFD seed virus was injected intra-cerebrally to five mice of 3–4 weeks old. Plain medium was injected intra-cerebrally to a group of five mice that served as control group and another five mice were injected with undiluted seed virus that served as positive control. Mice were observed for 14 days at 12-hour intervals for disease symptoms. At the end of 14 days, virus dilutions of 10⁻2, 10⁻3, 10⁻4, 10⁻5 and 10⁻6 caused disease symptoms in 100% of mice, whereas virus dilution of 10⁻7 caused disease in 20% of mice and virus dilution of 10⁻8 did not cause symptoms of the disease in any mice. The control group of mice that received plain medium remained healthy during the study period. All the mice that received undiluted virus showed classical symptoms of KFD.</p