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

Not Available

Not AvailableBrucellosis still remains an infectious, highly contagious and re-emerging endemic zoonosis especially in the Mediterranean and Middle-East regions of the world involving many countries including India where it constitutes occupational hazard (Shakerian et al., 2013 and Khamesipour et al., 2014). It also poses a serious threat to livestock economy by causing abortion, loss of offspring, infertility and reduction in milk yield. The prevalence of brucellosis in animal reservoirs is an evidence of its prevalence in human population and control of animal brucellosis is the key to its control in humans. Early diagnosis is essential to minimise the spread of the disease besides public health importance. In the present study molecular characterization of five Brucella melitensis isolates was carried out through Bruce-ladder multiplex PCR and compared with Brucella abortus, Brucella melitensis and Brucella suis vaccine and challenge strains. The amplification profile confirmed the isolates as Brucella melitensis and there was a significant difference among these field isolates with that of reference vaccine strains and the amplicons of all the field isolates were similar to amplicons of reference challenge strain, Brucella melitensis 16MNot Availabl

Not Available

Not AvailableIn this study, 225 milk samples were collected sequentially during 1st to 88th day from 25 HF cross cows in an organized farm. First five collections were obtained at a weekly interval (1,7,14,21 and 28 days) and later, fortnightly for two months (43, 58, 73 and 88 days). These milk samples were screened for Subclinical mastitis (SCM) by Somatic Cell Count (SCC). Further, multiplex-PCR for detection of S.aureus, E.coli, S.agalactiae, S.dysgalactiae and S.uberis was employed to detect the major bacterial pathogens. The SCM positivity was assessed based on criteria of SCC ≥ 500,000 cells /ml. The study revealed the high prevalence of variable SCM pattern in milking cows by SCC (73.33 %) in sequentially collected milk samples over a period of 88 days. No specific pattern of prevalence of SCM was observed during the study period. The prevalence of SCM was not influenced by the stage of lactation. In all the stages of lactation and age groups S. aureus, Streptococci and E.coli were detected with the predominance of S. aureus. The varied distribution of organisms in different stages of lactation did not influence the prevalence of SCM. Further, the high prevalence of SCM was noticed in aged cows. Among these, maximum number of milk samples (46 %, 52/113) revealing the presence of pathogens were obtained from cows in the age group 7-11 years. The multiplex PCR was found an easy and rapid method to detect the predominant pathogens causing SCM. The findings emphasize the need to control SCM through sequential monitoring of SCM through SCC, multiplex-PCR and proper managemental practices.Not Availabl

Not Available

Not AvailableSheep and goat brucellosis caused by Brucella melitensis, one of the most virulent Brucella species accounting for economic losses through abortion, stillbirths, reduction of milk yield and infertility. Disease has wide socioeconomic impact, in countries where, livestock sector is the major source of rural income. Early diagnosis is essential to minimise the spread of the disease besides public health importance. The present study reports the isolation, identification, biotyping and molecular confirmation of Brucella spp. in 18 different sheep and goat farms in Karnataka suspected to have brucellosis. A total of 550 serum samples, 25 aborted foetuses, uterine discharges and placental tissues were collected. The serum samples were subjected to Rose Bengal Plate Test (RBPT) and Competitive ELISA (c-ELISA). The clinical samples were processed for cultural isolation on Brucella Agar Media with selective antibiotic supplements. A total of 200 (36.36%) and 260 (47.27 %) serum samples were positive by RBPT and c-ELISA, respectively, further 195 (35.45 %) of them being positive by both the tests. Five Brucella isolates were recovered from 100 clinical samples. The isolates were characterized to their species by growing them on Brucella specific medium, biochemical reactions, CO2 requirement, H2S production, agglutination with A and M mono-specific antiserum, dye sensitivity to basic fuchsin and thionin. Further, molecular confirmation of the isolates was done by amplification of B. melitensis 16S (rRNA) sequence analysis by genus specific PCR and species specific IS711 repetitive DNA fragment by Brucella AMOS PCR. The present study envisages seroprevalence of at least 35.45 per cent and isolation rate of 25 per cent for B. melitensis warranting the need for institution of strict control measures.Not Availabl

Not Available

Not AvailableThe present study reports the epidemiological and virological investigations of the twelve different outbreaks of Peste des Petits Ruminants (PPR) in goats and sheep flocks with considerable morbidity and mortality recorded from different places of Karnataka state in India during 2014-2015. Clinical samples were collected from the affected flocks or villages for laboratory investigation along with epidemiological parameters. The PPR virus (PPRV) antigen and its genome were detected in the infected tissues or swab materials by sandwich enzyme-linked immunosorbent assay (s-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The significant epidemiological parameters recorded include young animals being severely affected than adult animals, which showed symptoms suggestive of PPR and changing pattern of disease in term of severity of gross lesions and clinical signs. The source of infection was traced back to introduction of new animals from other flocks or from other states. Despite regular vaccination of sheep and goats in the state, under National Control Programme on PPR (NCP-PPR), outbreaks need to be carefully monitored due mainly to production losses in small ruminants.Not Availabl

Not Available

Not AvailableA study was conducted to understand the current scenario of Classical swine fever (CSF) disease in Karnataka. Serum samples were collected from 517 pigs from 20 different districts of Karnataka and were tested for the presence of CSF antibodies. The prevalence of CSF antibodies from serum samples for the whole of Karnataka was 33% (173/517) in 20 districts. The southern Karnataka has the highest prevalence, which confirms the endemicity of the disease in that region and lowest prevalence in the northern Karnataka region. This is the first report that describes the seroepidemiology of CSF from Karnataka.Not Availabl

Reorientation Dynamics and Micromanipulation of Natural Microscopic Soft Matter in an Optical Trap with Varying Polarization of the Laser

We report here on the reorientation dynamics of the avian Red Blood Cell (aRBC), a structure ellipsoidal in shape and with a non-uniform distribution of birefringence across its diameter. We find that in linearly polarized light, an aRBC shows a dual reorientation behavior with the first reorientation about the major axis and the second about the minor axis so as to align its major axis along the laser propagation direction. We are able to explain the observed sequence of reorientation as a consequence of the minimization of work done in rotating the avian cell first about the major axis and then about the minor axis. These calculations are also used to also show why a single reorientation about the minor axis to reach the final equilibrium position is not seen experimentally. Further, in the case of elliptically polarized light, we see that the second reorientation process about the minor axis of the cell can be controlled by the ellipticity of the polarization of light. We explain this as a consequence of balance between the torques due to birefringence and reorientation

Not Available

Not AvailableIsolation, serological tests and molecular methods were adopted for the diagnosis of brucellosis in sheep flocks with history of reproductive disorders. Three serological tests RBPT, STAT and iELISA were employed to screen 527 sheep from 6 farms. Among serological tests, iELISA detected higher positives (14.80%), than RBPT (5.88%) and STAT (4.17%). Brucella melitensis was isolated from 6 of 33 seropositive sheep and none from seronegative animals. All the isolates were identified as B. melitensis by biochemical tests and confirmed by genus and species specific - AMOS PCR. None of the isolates yielded products of 285bp, 976bp and 498bp specific for B. suis, B. ovis and B. abortus, respectively. Of the 33 seropositive sheep, the bcsp31genus specific amplicon of 223bp was observed in 26 vaginal, 6 serum and 2 in blood samples. Vaginal samples was found suitable clinical sample for the isolation and PCR, followed by serum and blood. Different diagnostic approaches used in the present study are of value as anti Brucella antibodies and antigen were detected confirming the disease in sheep flock. This practically oriented approach is of immense importance, as it enables to institute the control strategies at the earliest.Not Availabl

<span style="font-size:13.0pt;font-family: "Times New Roman";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN-GB">Rapid typing of <i style="mso-bidi-font-style:normal">Pasteurella multocida</i> isolates from haemorrhagic septicaemia cases by multiplex PCR for their virulence</span>

124-126<span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"times="" roman";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">A total of 12 P. multocida isolates recovered from field outbreaks of haemorrhagic septicaemia in buffaloes were studied for the presence of 4 virulence genes (pfhA, tbpA, <i style="mso-bidi-font-style: normal">hgbB & toxA) in comparison with vaccine strain by multiplex PCR. The study showed the presence of pfhA and tbpA in all the field isolates and vaccine strain. None of the isolates harboured <i style="mso-bidi-font-style: normal">toxA and hgbB. The virulence genes tbPA and pfhA appear to be closely related to bovine P. multocida serogroup B isolates and are important for severe form of the disease. </span

Antimicrobials Influence Bond Stiffness and Detachment of Oral Bacteria

Oral biofilm can never be fully removed by oral hygiene measures. Biofilm left behind after brushing is often left behind on the same sites and exposed multiple times to antimicrobials from toothpastes and mouthrinses, after which removal becomes increasingly difficult. On the basis of this observation, we hypothesize that oral bacteria adhering to salivary conditioning films become more difficult to remove after adsorption of antimicrobials due to stiffening of their adhesive bond. To verify this hypothesis, bacteria adhering to bare and saliva-coated glass were exposed to 3 different mouthrinses, containing chlorhexidine-digluconate, cetylpyridinium-chloride, or amine-fluoride, after which bacterial vibration spectroscopy was carried out or a liquid-air interface was passed over the adhering bacteria to stimulate their detachment. Brownian motion-induced nanoscopic vibration amplitudes of 4 oral streptococcal strains, reflecting their bond stiffness, decreased after exposure to mouthrinses. Concurrently, the percentage detachment of adhering bacteria upon the passage of a liquid-air interface decreased after exposure to mouthrinses. A buffer control left both vibration amplitudes and detachment percentages unaffected. Exposure to either of the selected mouthrinses yielded more positively charged bacteria by particulate microelectrophoresis, suggesting antimicrobial adsorption to bacterial cell surface components. To rule out that exposure of adhering bacteria to the mouthrinses stimulated polysaccharide production with an impact on their detachment, Fourier transform infrared spectroscopy was carried out on bacteria adhering to an internal reflection element, prior to and after exposure to the mouthrinses. Infrared absorption band areas indicated no significant change in amount of polysaccharides after exposure of adhering bacteria to mouthrinses, but wave number shifts demonstrated stiffening of polysaccharides in the bond, as a result of antimicrobial adsorption to the bacterial cell surface and in line with changes in surface charge. Clinically, these findings suggest that accumulation of oral biofilm exposed to antimicrobials should be prevented (interdental cleaning aids, floss use), as removal becomes progressively more difficult upon multiple exposures