Evaluation of DNA Extraction Methods of Mule Dung

DNA isolation is a critical step in microbial community analysis of animal dung. DNA isolation from mule dung is challenging due to microbial diversity, composition and chemical nature of mule dung. Therefore, selection of an appropriate DNA isolation method is important to analyse the complete microbial diversity. In the current study, we evaluated the DNA isolation from mule dung samples (n=11) using QiAmp Mini stool kit as per manufacturer’s procedure with modifications. The results suggest that modifications in proprietary column based method improved the DNA quality and quantity suitable for mule dung microbial community analyses

Applications of Personalised Phage Therapy highlighting the importance of Bacteriophage Banks against Emerging Antimicrobial Resistance

Emerging antibiotic resistance is one of the most important microbiological issues of the 21st century. This poses a query regarding the future use of antibiotics and availability of other promising therapeutic alternatives. The awareness about antibiotic misuse has improved insufficiently and is evident by the increased incidences of multidrug resistant infections globally. Amongst different antibacterial therapeutic approaches phage therapy has created a niche of its own due to continuous use for treatment of human infections in Eastern Europe. Synergistic compounds along with phages have also been proposed as a better alternative compared to antibiotics or phage alone for treatment of chronic cases and seriously debilitating diseases. As such, why not allow custom made phage therapy for treatment of chronic infections? However, the success of phage therapy will depend upon instant availability of characterised bacteriophages from bacteriophage banks which may serve as the major catalyst in bringing Phage Therapy to main stream treatment alternatives or in combination therapy at least. In the current article we present a glimpse of comprehensive approach about utility of bacteriophage banks and further present personalised phage therapy in a synergistic role with antibiotics to overcome emerging antimicrobial resistance

A Study on the Effect of Laser Surface Texturing on Friction and Wear Behavior of Graphite Cast Iron

Dimples with various pitches and densities were produced using laser surface texturing (LST) to improve the friction and wear behavior of graphite cast iron. The objective of this study is to investigate the effectiveness of dimples on the friction and wear behavior of an internal combustion engine (ICE) cylinder. The specimens with a dimple pitch of 150 lm and a dimple density of 13% exhibited the lowest friction coefficient among the specimens, while the specimens with a dimple pitch of 200 lm and a density of 7% exhibited the highest resistance to wear

Dopunska prehrana ribljim uljem poboljšala je funkciju jajnika, koncepciju i određene reprodukcijske pokazatelje kod kobila pasmine marvari

We investigated the effect of dietary fish oil supplementation on the development of the ovarian follicles, corpus luteum (CL), conceptus and certain reproductive events in Marwari mares, since it is reported to improve reproduction in cows. Accordingly, non-lactating mares (n = 20) were randomly assigned into two groups (10 per group) and fed either the control diet (CTR) or a diet enriched with fish oil (FOS) to supplement n-3 polyunsaturated fatty acids (PUFA) at the rate of 64 mg/kg body weight/day for 70 days or until 45 days post-ovulation in the mares that became pregnant. Estrus was detected using a teaser and insemination was performed using frozen thawed semen in the experimental mares. Development of the ovarian follicle, CL and conceptus were recorded using trans-rectal ultrasonography. Plasma concentrations of progesterone and estradiol were estimated by radioimmunoassay. In the FOS group, the diameter of the largest follicle from day 4 of estrus until ovulation, and the diameter of the CL on day 7 post-ovulation (D7PO) were greater (P<0.05). However, on day 15 post-ovulation (D15PO), the CL diameter increased significantly in the pregnant mares. Dietary fish oil significantly improved the development of the embryo as evidenced by an increase in the diameter of the embryonic vesicle on day 15 post-ovulation (D15PO), and the embryo proper on day 28 post ovulation (D28PO). Further, the mean plasma estradiol concentration was higher on the day of estrus onset (P<0.05) and day 4 of estrus (P<0.01) in the FOS group. Similarly, dietary fish oil significantly increased the plasma progesterone on D15PO in the pregnant mares (P<0.01). Although the duration of estrus was shorter by 19 hours (P<0.05), the length of the estrous cycle did not vary in the FOS group. A non-significant increase in the pregnancy rate was observed in the mares that received fish oil. It was concluded that dietary fish oil supplementation improved ovarian function and embryonic development in the Marwari mares.Na temelju prethodnih izvješća o poboljšanju reprodukcije krava, kod kobila pasmine marvari istražili smo učinak ribljeg ulja kao prehrambenog dodatka na razvoj folikula jajnika, žutog tijela, koncepcije i određenih reprodukcijskih pokazatelja. U skladu s ciljem, 20 kobila koje nisu bile u laktaciji je metodom slučajnog izbora podijeljeno u dvije skupine s po 10 kobila. Za razliku od kontrolne skupine, kobile u eksperimentalnoj skupini hranjene su obrokom obogaćenim ribljim uljem u obliku dodatka koji je sadržavao n-3 polinezasićene masne kiseline (PUFA), u dnevnoj količini od 64 mg na jedan kg tjelesne mase. Eksperiment je trajao 70 dana, odnosno za kobile koje su ostale gravidne do 45 dana nakon ovulacije. Estrus je praćen teaserom, a osjemenjivanje je provedeno zamrznutim sjemenom. Razvoj folikula jajnika i žutih tijela te gravidnosti praćeni su transrektalnim ultrazvukom. Koncentracija progesterona i estradiola u plazmi procijenjena je radioimunološkom metodom. U eksperimentalnoj skupini utvrđen je veći (P<0,05) promjer najvećeg folikula od 4. dana estrusa do ovulacije i veći promjer žutog tijela 7. dan nakon ovulacije. No, 15. dan nakon ovulacije promjer žutih tijela bio je signifikantno povećan kod gravidnih kobila. Obrok obogaćen ribljim uljem signifikantno je poboljšao razvoj embrija, što se očitovalo povećanjem promjera embrionalne vezikule 15. dan nakon ovulacije i povećanjem embrija 28. dan nakon ovulacije. Nadalje, u eksperimentalnoj skupini kobila srednja koncentracija estradiola u plazmi bila je veća na dan početka estrusa (P<0,05) i 4. dan estrusa (P<0,01). Slično tome, obrok obogaćen ribljim uljem kod gravidnih je kobila signifikantno (P<0,01) povećao progesteron u plazmi 15. dan nakon ovulacije. U kobila eksperimentalne skupine je, uz varijacije estrusnog ciklusa, utvrđeno i za 19 sati kraće trajanje estrusa (P<0,05). Povećana stopa gravidnosti kod kobila dohranjivanih ribljim uljem nije bila signifikantna. Zaključeno je da je hranidba s dodatkom ribljeg ulja kod kobila pasmine marvari poboljšala funkciju jajnika i razvoj embrija

Inhibitor of Sarco/Endoplasmic Reticulum Calcium-ATPase Impairs Multiple Steps of Paramyxovirus Replication

Sarco/endoplasmic reticulum calcium-ATPase (SERCA) is a membrane-bound cytosolic enzyme which is known to regulate the uptake of calcium into the sarco/endoplasmic reticulum. Herein, we demonstrate for the first time that SERCA can also regulate virus replication. Treatment of Vero cells with SERCA-specific inhibitor (Thapsigargin) at a concentration that is nontoxic to the cells significantly reduced Peste des petits ruminants virus (PPRV) and Newcastle disease virus (NDV) replication. Conversely, overexpression of SERCA rescued the inhibitory effect of Thapsigargin on virus replication. PPRV and NDV infection induced SERCA expression in Vero cells, which could be blocked by Thapsigargin. Besides inducing enhanced formation of cytoplasmic foci, Thapsigargin was shown to block viral entry into the target cells as well as synthesis of viral proteins. Furthermore, NDV was shown to acquire significant resistance to Thapsigargin upon long-term passage (P) in Vero cells. As compared to the P0 and P70-Control, the fusion (F) protein of P70-Thapsigargin virus exhibited a unique mutation at amino acid residue 104 (E104K), whereas no Thapsigargin-associated mutations were observed in HN gene. To the best of our knowledge, this is the first report describing the virus-supportive role of SERCA and a rare report suggesting that viruses may acquire resistance even in the presence of an inhibitor that targets a cellular factor

A Comprehensive Review on Equine Influenza Virus:Etiology, Epidemiology, Pathobiology, Advances in Developing Diagnostics, Vaccines, and Control Strategies

Among all the emerging and re-emerging animal diseases, influenza group is the prototype member associated with severe respiratory infections in wide host species. Wherein, Equine influenza (EI) is the main cause of respiratory illness in equines across globe and is caused by equine influenza A virus (EIV-A) which has impacted the equine industry internationally due to high morbidity and marginal morality. The virus transmits easily by direct contact and inhalation making its spread global and leaving only limited areas untouched. Hitherto reports confirm that this virus crosses the species barriers and found to affect canines and few other animal species (cat and camel). EIV is continuously evolving with changes at the amino acid level wreaking the control program a tedious task. Until now, no natural EI origin infections have been reported explicitly in humans. Recent advances in the diagnostics have led to efficient surveillance and rapid detection of EIV infections at the onset of outbreaks. Incessant surveillance programs will aid in opting a better control strategy for this virus by updating the circulating vaccine strains. Recurrent vaccination failures against this virus due to antigenic drift and shift have been disappointing, however better understanding of the virus pathogenesis would make it easier to design effective vaccines predominantly targeting the conserved epitopes (HA glycoprotein). Additionally, the cold adapted and canarypox vectored vaccines are proving effective in ceasing the severity of disease. Furthermore, better understanding of its genetics and molecular biology will help in estimating the rate of evolution and occurrence of pandemics in future. Here, we highlight the advances occurred in understanding the etiology, epidemiology and pathobiology of EIV and a special focus is on designing and developing effective diagnostics, vaccines and control strategies for mitigating the emerging menace by EIV

Persons ‘never treated’ in mass drug administration for lymphatic filariasis: identifying programmatic and research needs from a series of research review meetings 2020–2021

As neglected tropical disease programs rely on participation in rounds of mass drug administration (MDA), there is concern that individuals who have never been treated could contribute to ongoing transmission, posing a barrier to elimination. Previous research has suggested that the size and characteristics of the never-treated population may be important but have not been sufficiently explored. To address this critical knowledge gap, four meetings were held from December 2020 to May 2021 to compile expert knowledge on never treatment in lymphatic filariasis (LF) MDA programs. The meetings explored four questions: the number and proportion of people never treated, their sociodemographic characteristics, their infection status and the reasons why they were not treated. Meeting discussions noted key issues requiring further exploration, including how to standardize measurement of the never treated, adapt and use existing tools to capture never-treated data and ensure representation of never-treated people in data collection. Recognizing that patterns of never treatment are situation specific, participants noted measurement should be quick, inexpensive and focused on local solutions. Furthermore, programs should use existing data to generate mathematical models to understand what levels of never treatment may compromise LF elimination goals or trigger programmatic action

A community based interventional approach to intranatal And neonatal health care

Objective : To assess the contribution of the interventions through the local change agent (Bal Parivar Mitra) towards achievement of health awareness among rural women regarding maternal child health through improvement and change in health practices. Study design: Stratified multistage random sampling technique. Study unit : Within the selected households lactating mothers were selected as study units. Study area : The study was conducted in Jasra and Saidabad blocks of Allahabad district wherein MCHN Project is going on since July 2000. Study variable : Maternal care. Intervention, Impact. Statistical analysis : Ztesl for testing significance of differences between two proportions (Z - test). Results: Deliveries assisted by trained persons increased from 22.4%to 36.7%. Follow-up of'5-cleans’ during pregnancy was among 43.3%. Birth registration increased from 19.2% in baseline to 35%. feeding of colostrum from 27,4% to 40.0%. Breastfeeding within half an hour after birth was among 23.3% followed by 16.7% within 1/2-12 hours. Proper warmth was given to 68.3% newborns and 58.3% babies were bathed after one day of birth. Conclusions : The suggested intervention package through BPM seems to be a sustainable effort and several parameters of intranatal and neonatal health care arc expected to be attained as long-term achievements

Comparative evaluation of diagnostic tests for the detection of Mycobacterium avium subsp. paratuberculosis in the tissues of sheep affected with distinct pathology of paratuberculosis

Aims and objective: Paratuberculosis or Johne's disease is a chronic infectious granulomatous enteritis, mainly of cattle, sheep, goats, and other domestic and wild animals caused by Mycobacterium avium subsp. paratuberculosis (MAP). Currently, MAP has been recognized as an important animal pathogen with significant zoonotic and public health concerns. The early detection of infected animals using suitable diagnostic methods helps in developing control and preventive strategies for the herd. Therefore, the present study was aimed to determine the comparative efficacy of certain diagnostic methods used in the identification and confirmation of MAP in the ovine tissues with distinct pathology of paratuberculosis. Methods: The ileum and mesenteric lymph node (MLN) tissues were collected from 38 sheep infected with paratuberculosis from organized farms of Rajasthan. These animals were further classified as paucibacillary (PB; n = 15) or multibacillary (MB; n = 23) on the basis of histopathological findings and mycobacterial loads. The ileum and MLN tissues of these animals were subjected to IS900 and 251 gene polymerase chain reaction (PCR) and bacterial culture. The tissue sections from MB, PB, and uninfected control sheep groups were stained using indirect immunoperoxidase technique (IPT) and Ziehl–Neelsen (ZN) method. Results: On bacterial culture examination of the ileum and MLN tissues using Herrold's egg yolk medium, MAP was isolated in 14 (60.9%) MB and 5 (33.3%) PB sheep. Of 38 sheep, IS900 PCR detected 21 (55.2%) positive for MAP, of which 19 (82.6%) were MB and 2 (13.3%) were PB sheep. Similarly, 251 gene PCR detected 25 (65.7%) sheep positive for MAP infection, of which 21 (91.3%) were MB and 4 (27%) were PB sheep. Thus, 251 gene PCR was found superior to IS900 PCR in the detection of MAP from the tissues. In PB sheep, IPT and ZN tests were positive in 87.5% and 50% of ileum sections and 70% and 37.5% in MLN sections, respectively. In MB sheep, IPT and ZN tests detected all animals as positive for MAP organisms or antigen and had equal sensitivity in the detection of MAP. The overall sensitivity of IPT was found superior (95%) to ZN staining (80%) in the demonstration of acid-fast bacteria or its antigen in the tissues. Conclusion: The sensitivity of all the tests in the detection of MAP was lower in PB sheep than in MB sheep. Bacterial culture detected MAP in only 50% of sheep and was found less sensitive than other tests used in the present study. Comparing the overall sensitivity of both the PCR assays, 251 gene PCR was found superior to IS900 gene PCR. The sensitivity of IPT was found superior (95%) to the ZN staining (80%) in the demonstration of acid-fast bacteria in the tissues. In the present study, IPT was found superior in the detection of MAP in PB and MB form of ovine paratuberculosis. This test can be used in the confirmation of post mortem diagnosis and research of paratuberculosis along with histopathology. However, 251 gene PCR assay was found easier to perform than IPT and could be used as paratuberculosis screening test in endemic sheep farms for blood and fecal samples