Characterization of infectious laryngotracheitis virus isolates from laying hens during 2019-2020 outbreaks in Tamil Nadu, India.

Infectious laryngotracheitis (ILT) is an acute respiratory disease in chickens that is a serious threat to poultry-producing countries worldwide. In the present study, we isolated and characterized infectious laryngotracheitis (ILTV) virus isolates by sequencing and restriction fragment length polymorphism analysis of PCR-amplified products (PCR-RFLP). A total of 26 ILTV outbreaks were investigated that occurred between 2019 and 2020 in flocks that had not been vaccinated against ILTV. ILTV was isolated by cultivating tracheal samples in embryonated chicken eggs, which showed multiple opaque pock lesions and thickening of the chorioallantoic membrane after 120 hours of infection. The ILTV isolates were identified and characterized by PCR and sequencing a portion of the ICP4 and TK genes. Phylogenetic analysis based on the ICP4 region showed that the sequences clustered with chicken-embryo-origin vaccine-like strains. Sequence analysis of the ICP4 region differentiated chicken-embryo-origin (CEO), tissue-culture-origin (TCO), and field ILTV strains, with significant differences in nucleotide and amino acid sequences. Furthermore, PCR-RFLP analysis of the TK gene showed that the patterns were identical to those obtained with low-virulence and vaccine strains. In conclusion, sequencing of a portion of the ICP4 region of ILTV allowed differentiation of ILTV field, CEO, and TCO vaccine strains. In this study, CEO-vaccine-like strains were found to be the cause of ILTV outbreaks between 2019 and 2020 in Tamil Nadu in southern India.

Beak and bumper - Physics forceps: Evaluation of new technique in extraction.

PURPOSE: The history of dentistry is not short; it started from ancient Egypt to ancient Romans and ancient Greeks. When it comes to extraction, all of them have made their own discoveries and progress. The progress they made also helped dentistry to move ahead in evolution of new extraction technique - the Physics Forceps. We have assessed the viability in using the Physics Forceps for routine dental extraction in our study for a period of 1 year. MATERIALS AND METHODS: The study was conducted on 241 patients indicated for extraction based on our inclusion criteria using Physics Forceps after obtaining informed consent and University Ethics Committee approval. Tooth fracture, buccal alveolar bone fracture, and soft tissue injury around the tooth to be extracted were studied. RESULTS: In our present study of 241 patients, 57.67% were females and 42.32% were males, out of which 93.77% had no tooth fracture, 3.32% had crown fracture, 1.65% had root fracture, and 1.24% had apex fracture. Further, 85.47% had no buccal alveolar bone fracture and 14.53% had buccal alveolar bone fracture. Using proper technique, 96.26% of patients had no soft tissue damage, and minimal damage was seen in 3.73% of patients. DISCUSSION: Extraction by Physics Forceps is a very good technique of extraction. No or very minimal tooth fracture and soft tissue injury were noted. Though the forceps is costly, it represents a valuable addition to regular armamentarium for a general dentist for routine extraction. Physics Forceps is a dental extractor rather than a forceps.

Conventional and molecular determination of drug resistance in Mycobacterium tuberculosis and Mycobacterium bovis isolates in cattle.

This study was aimed out to explore the presence of drug resistance among M. tuberculosis and M. bovis isolates (n = 51) of bovine origin by conventional broth microdilution method and molecular methods. By broth microdilution method, 16 isolates were found to be resistant to isoniazid, 08 isolates were resistant to pyrazinamide, 09 isolates were resistant to rifampicin and 07 isolates were found to be resistant for ethambutol. Two isolates showed resistance to rifampicin and pyrazinamide, one isolate showed resistance to pyrazinamide and ethambutol and 03 isolates showed resistance to isoniazid and ethambutol. None of isolates showed multi drug resistance (MDR). Other than the M. bovis strains, none of the other M. tuberculosis isolates showed any resistance to pyrazinamide. Molecular methods by multiplex PCR targeting katG, pncA, rpoB genes, multiplex allele specific PCR to detect mutation in embB codon 306 and sequencing showed point mutation in katG and rpoB gene. No mutation could be detected in the embB gene by multiplex allele specific PCR. The results indicates that further elaborate studies need to be carried out due to the presence of drug resistant M. tuberculosis in bovines which could be due to spill over from human in tuberculosis endemic areas making TB eradication programme more challenging.

Sequence analysis of VP2 hypervariable region of the field isolates of infectious bursal disease viruses from southern region of India.

Infectious bursal disease virus isolates obtained from southern parts of India were subjected to comparative sequencing and phylogenetic analysis of 743bp hypervariable region of VP2. The sequence analysis showed that among eight isolates, only HY12 showed the characteristic conserved amino acid residues at 256I, 294I, and 299S of vvIBDV. Six isolates BGE14, PY12, NKL14, VCN14, RPM14 and EDE14 had conserved amino acid residues at 256I and 299S, whereas at residue 294, isoleucine was substituted by valine. The remaining isolate MB11 had leucine at residue 294 and asparagine at residue 299 similar to classical strain 52/70. The serine-rich heptapeptide sequence SWSASGS adjacent to the second hydrophilic region was conserved in all seven Indian IBDV isolates except isolate MB11. Conservation of this sequence was earlier reported to be an indication of a virus isolate being pathogenic in nature. The reported heptapeptide sequence of the classical strain is 'SWSARGS'. In the present study, 'SWSARGS' heptapeptide sequence was observed in MB11 isolate. The pathogenicity trials conducted with these isolates further confirmed the genome analysis in classification. This study further reveals that the circulating IBDV strains in India could be diverse in nature.

Molecular identification of Mycobacterium tuberculosis in cattle.

Bovine tuberculosis continued to be a re-emerging problem in some countries especially in endemic areas due to the fact that human and animal health surveillance system is not adopted to diagnose the infection. This crisis can be attributed due to sharing of the same habitat especially in rural areas. In the present study, a total of 148 samples were collected from cattle for isolation over a period of 3 years from cattle with and without lesions, of which 67 isolates were obtained by culture. Fifty one isolates were identified as Mycobacterium tuberculosis complex (MTBC) by IS6110 PCR of which 43 (84.3%) were identified as M. tuberculosis and 08 (15.6%) were identified as M. bovis by using 12.7kb fragment multiplex PCR. Among this, 31 isolates which were positive for IS6110 PCR were subjected to spoligotyping and revealed 28 isolates belonging to MANU1 strain of M. tuberculosis. This study clearly indicates that high prevalence of M. tuberculosis than M. bovis in bovine was identified by means of culture and by molecular methods M. tuberculosis can affect cattle producing lesion in contradiction to the earlier thoughts. This study speculates that M. tuberculosis MANU1 strain infection in cattle could be due to spill over from human or other non specific hosts in tuberculosis endemic areas. Though bovine tuberculosis due to M. tuberculosis in cattle is not considered a serious threat worldwide, in countries where human TB is endemic, M. tuberculosis infection of cattle needs to be considered.

Complete Genome Sequence Analysis of a Naturally Reassorted Infectious Bursal Disease Virus from India.

The novel infectious bursal disease virus (IBDV) isolate BGE14/ABT1/MVC/India is a very virulent IBDV that was isolated from broiler flocks in southern parts of India during 2014. Here, we report, for the first time in India, the complete genome sequence of BGE14/ABT1/MVC/India, a reassortment strain with segments A and B derived from a very virulent IBDV strain and an attenuated IBDV, respectively. The findings from this study provide additional insight into the genetic exchange between attenuated and very virulent strains of IBDV circulating in the field.

Complete Genome Sequences of Lineage IV Peste des Petits Ruminants Viruses from the Indian Subcontinent.

The complete genome sequences of two virulent lineage IV peste des petits ruminants viruses (PPRVs) isolated from clinically infected goats in the Indian subcontinent are reported here. This is the first report of a complete genome sequence of a virulent PPRV isolate from India in recent decades.

Fate of Free Fat Dermis Graft in TMJ Interpositional Gap Arthroplasty: A Long Term MRI Study.

INTRODUCTION: Free fat dermis graft is a good interpositional material for TMJ gap arthroplasty. Analysing the fate of the graft by magnetic resonance imaging (MRI) images helps in excellent visualization of both bony and soft tissue anatomy of the operated TMJ joint as well as in assessing the changes in dermis graft which was previously placed. PURPOSE OF THIS STUDY: To investigate the radiological fate of the dermis-fat graft within the TMJ using MRI. MATERIALS AND METHODS: Five joints of five patients who had dermis-fat grafts placed in their TMJ following gap arthroplasty were recruited for this study. Each patient had undergone TMJ gap arthroplasty with immediate dermis-fat graft placement. All the patients are then subjected to MRI. RESULTS: Fat graft was identified in close proximity to the mandibular condyle in all cases, with only three joints demonstrating fat covering the entire articular surface of the mandibular condyle. In the remaining joints the interpositional material found in the MRI defined joint space with mainly grey appearance, suggesting tissue change to other than fat, i.e. scar or granulation tissue. CONCLUSION: When free fat dermis graft is placed as interpositional material the graft occupied the entire TM joint and prevented it from recurrent ankylosis. The graft placed aids in normal functioning of the temperomandibular joint without any complications.

Rapid Detection of Avian Infectious Bronchitis Virus by Reverse Transcriptase-Loop Mediated Isothermal Amplification.

A reverse-transcription loop mediated isothermal amplification (RT-LAMP) was developed for rapid diagnosis of infectious bronchitis (IB) in poultry by targeting the spike protein 2 gene (S2). RT-LAMP primers were designed for IBV-S2 targets and optimized to run at 60 °C for 45 min. As compared with RT-PCR, RT-LAMP was 100 times more sensitive for IBV-S2 gene. RT-LAMP showed specific amplification with IB viral genome but not with other avian respiratory pathogens due to their mismatching with IBV-S2-RT-LAMP primers. RT-LAMP reaction products were visually detected by the addition of propidium iodide stain. Out of 102 field samples tested for detection of IBV, RT-LAMP detected IBV in 12 samples for S2 gene whereas RT-PCR detected IBV in six samples for S2 gene. The sensitivity of the RT-LAMP was 100 % and the specificity was 94 % for S2 gene. Since the developed RT-LAMP to detect IBV is simple, rapid, sensitive and specific, it can be a useful diagnostic tool for detection of IB in poultry in less equipped laboratories and in field conditions.

Molecular characterisation of lineage IV peste des petits ruminants virus using multi gene sequence data.

Peste des petits ruminants is responsible for an economically important plague of small ruminants that is endemic across much of the developing world. Here we describe the detection and characterisation of a PPR virus from a recent outbreak in Tamil Nadu, India. We demonstrate the isolation of PPR virus from rectal swab and highlight the potential spread of disease to in-contact animals through faecal materials and use of faecal material as non-invasive method of sampling for susceptible wild ruminants. Finally we have performed a comprehensive 'multi-gene' assessment of lineage IV isolates of PPRV utilising sequence data from our study and publically available partial N, partial F and partial H gene data. We describe the effects of grouping PPRV isolates utilising different gene loci and conclude that the variable part of N gene at C terminus gives the best phylogenetic assessment of PPRV isolates with isolates generally clustering according to geographical isolation. This assessment highlights the importance of careful gene targeting with RT-PCR to enable thorough phylogenetic analysis.

Marker assisted selection of low phytic acid trait in maize (Zea mays L.).

Maize is the third important major food crop. Breeding for low phytate maize genotypes is an effective strategy for decreasing the content of kernel phytic acid (a chelator of cations such as Ca(2+) and Fe(3+) ) and thereby increasing the bioavailability of nutritive minerals in human diet and animal feed. Previous studies have established that a mutant plant with a lpa2-2 allele accumulates less phytic acid in seeds. Therefore, the marker assisted backcross breeding (MABB), which involves introgression of lpa2-2 recessive allele (which confer low phytate trait) from a lpa2-2 mutant line into a well-adapted line using backcrosses and selection of lines possessing lpa2-2 allele in each backcross population using molecular markers, is an effective strategy for developing low phytate maize. So far, no studies have developed any lpa2-2 allele specific molecular markers for this purpose. Here, using backcross and selfed progenies, obtained by crossing low phytate mutant line 'EC 659418' (i.e. donor of lpa2-2 allele) into agronomically superior line 'UMI395', we have validated that a SSR marker 'umc2230', located 0.4 cM downstream of lpa2-2, cosegregate, in a Mendelian fashion, with low phytic acid trait. Therefore umc2230 can be dependably used in MABB for the development of low phytate maize.

The versatility of lower eyelid blepharoplasty incision: In open reduction and fixation of zygomatic complex injuries.

INTRODUCTION: A ZYGOMATIC COMPLEX FRACTURE INCLUDES DISRUPTION OF THE FOUR ARTICULATING SUTURES: zygomaticofrontal, zygomaticotemporal, zygomaticomaxillary and zygomaticosphenoidal sutures. All zygomatic complex fractures involve the orbital floor and therefore an understanding of orbital anatomic features is essential for those treating these injuries. AIMS AND OBJECTIVES: To analyze the efficacy and shortcomings of this approach. To evaluate the adequacy, role of tarsorrhaphy, difficulties, role of steroid in postoperative edema control in lower lid blepharoplasty approach. MATERIALS AND METHODS: A total number of six patients were included in this study and all the patients were treated surgically under general anesthesia. All the patients were approached through lower eyelid blepharoplasty incision. The first skin crease in the lower eyelid region is selected for this incision. RESULTS: All patients were administered with steroid injection. Frost sutures were placed in four cases and tarsorrhaphy was done in two patients. Three cases encountered immediate mild edema and immediate scar formation. Late scar was present only in two patients with a follow up of three months. CONCLUSION: Lower eyelid blepharoplasty incision is an excellent, non complicated, simple procedure in the management of fractures in the infraorbital region, orbital floor, which occurs as a part of zygomatic complex fractures.

Calcium phosphate coupled Newcastle disease vaccine elicits humoral and cell mediated immune responses in chickens.

Calcium phosphate (CaP) particles were coupled with inactivated Newcastle disease virus (NDV) vaccine. The surface morphology of CaP particles coupled to NDV was found to be spherical, smooth and with a tendency to agglomerate. The mean (± SE) size of CaP particles was found 557.44 ± 18.62 nm. The mean percent encapsulation efficiency of CaP particles coupled to NDV assessed based on total protein content and haemagglutination (HA) activity in eluate was found to be 10.72 ± 0.89 and 12.50 ± 2.09, respectively. The humoral and cell mediated immune responses induced by CaP coupled NDV vaccine were assessed in comparison to a commercial live vaccine (RDV 'F'). CaP coupled NDV vaccine elicited prolonged haemagglutination inhibition (HI) and enzyme linked immunosorbent assay (ELISA) titres in the serum even at fourth and fifth week post-vaccination (PV), unlike RDV 'F' inoculated chickens whose titres declined to insignificant levels by this time. CaP coupled NDV vaccine could stimulate HI antibodies in tracheal washings and tears from second and first week PV, respectively. IgA ELISA antibodies were also seen in tracheal washings of these birds from third week PV and in tears from second week PV. CaP coupled NDV vaccine elicited cell mediated immune responses (CMI) from two to four weeks PV. The stimulation indices obtained after stimulation with specific antigen was not significantly different between CaP coupled antigen and live NDV virus except on first week PV. However, CaP coupled antigen did not cause suppression of lympo proliferation as indicated by statistically similar responses to mitogen, concanavalin A between the two groups. Overall, CaP coupled NDV vaccine elicited stronger and prolonged immune responses in comparison to the commercial live vaccine. No increase in the serum calcium and phosphorous levels were seen in CaP coupled NDV vaccine inoculated chickens.

Versatility of full thickness skin-subcutaneous fat grafts as interpositional material in the management of temporomandibular joint ankylosis.

The authors present a review of seven patients (eight joints) with temporomandibular ankylosis treated between 2007 and 2008. The aim of this retrospective study was to present the experience of using full thickness skin-subcutaneous fat grafts, harvested from the patient's abdomen as interpositional material after gap arthroplasty. All patients presented with osseous ankylosis and were graded according to Topazian's classification. Postoperative follow up ranged from 12 to 24 months. Maximal inter-incisal opening (MIO) on presentation ranged from 0 to 8mm, which stabilized to 27-44mm at follow up. There was no evidence of re-ankylosis. This study found merit in the use of autogenous full thickness skin-subcutaneous fat graft as an interpositional material for up to 2 years following ankylosis release.

Are people aware of oral and maxillofacial surgery in India?

INTRODUCTION: Maxillofacial surgery is one of the most rapidly expanding specialities in India. There is however a very poor understanding of the scope of the speciality. This paper attempts to find out the awareness of the speciality in India. MATERIALS AND METHODS: A questionnaire was given to 50 medical, dental undergraduate students, 50 medical and dental practitioners, and 50 lay persons giving us a total of 250 responses. 10 common anomalies were chosen and they were asked to choose the most appropriate speciality which they would ask to treat the problem. RESULTS: While trauma and facial deformity correction were recognised to be mainly treated by maxillofacial surgeons, the other maxillofacial problems were poorly recognised to be treated by our speciality. CONCLUSION: This study highlights the need to promote our speciality among the dentists, doctors and general public. and if need be even change the name of our speciality from oral and maxillofacial surgery to a more simple but more easily understood facial surgery.

Myositis ossificans traumatica of the medial pterygoid.

Myositis ossificans traumatica (MOT) is a non-neoplastic, heterotopic bone formation within muscle or fascia, presumably due to acute trauma, a burn, surgical manipulation, or repeated injury. A 36-year-old woman presented with progressive limitation of mouth opening over the past 5 months. Computed tomography revealed an irregular ossified mass attached to the medial aspect of the left ramus, extending to the medial and lateral pterygoid plates. MOT of the medial pterygoid is rare, with only four previously reported cases in the English language literature. Surgical excision of the ossification is the main treatment modality. The authors were able to establish a cleavage plane of 3 cm between the bony mass and the medial surface of the ramus. Abdominal fat was placed as an interpositional material to prevent fibrosis and heterotopic bone formation.

Comparative efficacy of peste des petits ruminants (PPR) vaccines.

Peste des petits ruminants (PPR) is a highly contagious, economically important viral disease of sheep and goats with high morbidity and mortality rates. In order to control the disease effectively, highly sensitive diagnostic tests coupled with potent vaccines are important pre-requisites. At present, there are three live attenuated PPR vaccines available in India including Sungri 96, Arasur 87 and Coimbatore 97. Indian Veterinary Research Institute (IVRI) Mukteswar developed the PPR Sungri 96 (isolate of goat origin) vaccine; while Tamil Nadu Veterinary and Animal Sciences University (TANUVAS) developed the Arasur 87 (isolate of sheep origin) and Coimbatore 97 (isolate of goat origin). In this study, the potency of these vaccines including a fourth vaccine from Institute of Animal Health and Veterinary Biologicals, Bangalore (IAH&VB) were tested as per the office International des Epizooties (OIE) guidelines by challenge studies in sheep and goats and their efficacies were evaluated using PPR C-ELISA. Potency tests of these vaccines in sheep and goats revealed that three of the vaccines were potent; however, the IAH &VB vaccine was comparatively less potent. The three vaccines could presumably be used for mass vaccination of both sheep and goats while contemplating PPR control program.

Impact of impacted mandibular third molars in mandibular angle and condylar fractures.

Previous retrospective analyses prove that impacted mandibular third molars (M3s) increase the risk of angle fractures and decrease the risk of concomitant fractures to the condyle. The authors have attempted to verify these relationships and identify the underlying mechanism of injury. A retrospective cohort was designed for patients attending the Division of Oral and Maxillofacial Surgery from January 2001 till October 2008. The primary predictor variable was M3. The secondary predictor variables were: M3 position, classified using the Pell and Gregory system; angulation, classified using Shiller's method; and the number of visible dental roots. The outcome variables were angle and condyle fractures. Hospital charts and radiographs were used to determine and classify these variables. The study sample comprised 1102 mandibular fractures in 600 patients. For patients injured by moderate traumatic force resulting in two fractures of the mandible, the presence/absence of impacted M3s played an important role in angle/condylar fractures. Patients with impacted M3s were three times more likely to develop angle fractures and less likely to develop condylar fractures than those without impacted M3s. This study provides clinical evidence to suggest that the removal of unerupted mandibular third molars predisposes the mandible to condyle fractures.

Expression profile of toll like receptors in a range of water buffalo tissues (Bubalus bubalis).

The present study was carried out to determine the expression profile of toll-like receptors (TLRs) 1-10 in buffalo peripheral blood mononuclear cells (PBMNC), neutrophils, spleen, liver, lung, heart, kidney, ovary and uterus using reverse transcriptase polymerase chain reaction (RT-PCR) with bovine TLR-specific primers The buffalo TLR partial nucleotide sequences had 95-98% nucleotide homology with bovine TLR sequences available in the GenBank. PBMNC expressed all TLRs except TLR1 and neutrophils expressed all TLRs except TLR3. Expression of all TLRs was observed in spleen, lung and liver tissues. Wide range of TLR mRNA expression was observed in heart, which lacked the expression of only TLR10. Among the tissues analyzed kidneys had the least repertoire of TLR expression. The kidney tissue revealed mRNA expression of only TLR2, TLR5, TLR7 and TLR9. Among the reproductive tissues analyzed, uterus expressed a wide range of TLRs such as 2, 5, 7, 8, 9 and 10 while ovary expressed all TLRs except TLR1 indicating their immuno competence.